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Volume 269
Handbook of Experimental
Pharmacology
Editor-in-Chief
James E. Barrett
Center for Substance Abuse Research, Lewis Katz School of Medicine at
Temple University, Philadelphia, PA, USA
Editorial Board
Veit Flockerzi
Institute for Experimental and Clinical Pharmacology and Toxicology,
Saarland University, Homburg, Germany
Michael A. Frohman
Center for Developmental Genetics, Stony Brook University, Stony Brook,
NY, USA
Pierangelo Geppetti
Headache Center, University of Florence, Florence, Italy
Franz B. Hofmann
Forschergruppe 923 Carvas, Technical University, München, Germany
Rohini Kuner
Institute of Pharmacology, Heidelberg University, Heidelberg, Germany
Martin C. Michel
Department of Pharmacology, University of Mainz, Mainz, Germany
Clive P. Page
SIPP, Kings College London, London, UK
KeWei Wang
School of Pharmacy, Qingdao University, Qingdao, China
The Handbook of Experimental Pharmacology is one of the most

authoritative and influential book series in pharmacology. It provides
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More information about this series at https://link.springer.com/
bookseries/164
Editors
Gunnar Schulte and Pawel Kozielewicz
Pharmacology of the WNT Signaling

System
1st ed. 2021
Editors
Gunnar Schulte
Department of Physiology & Pharmacology Section for Receptor
Biology & Signaling, Karolinska Institutet Biomedicum 6D, Stockholm,
Sweden
Pawel Kozielewicz
Department of Physiology & Pharmacology Section for Receptor
Biology & Signaling, Karolinska Institutet Biomedicum 6D, Stockholm,
Sweden
ISSN 0171-2004 e-ISSN 1865-0325

Handbook of Experimental Pharmacology
ISBN 978-3-030-85498-0 e-ISBN 978-3-030-85499-7
https://doi.org/10.1007/978-3-030-85499-7
© The Editor(s) (if applicable) and The Author(s), under exclusive

license to Springer Nature Switzerland AG 2021
This work is subject to copyright. All rights are solely and exclusively
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Preface
The WNT family of secreted lipoglycoproteins was discovered more
than 35 years ago and it is established as a key player in embryonic
development, tissue regeneration, and homeostasis as well as many
devastating disorders such as developmental defects, diverse forms of
tumors, bone disease, fibrosis, and neurodegenerative disorder (Willert
and Nusse 2012). The 19 mammalian WNTs interact with a complex
array of cell surface receptors, of which the Frizzleds (FZD1–10),
members of the Class F of G protein-coupled receptors (GPCRs),
present the main WNT receptors, which act either alone or in concert
with coreceptors such as LRP5/6, ROR1/2, RYK, and PTK7 (Schulte
2010; van Amerongen 2012).
While many of the players of the WNT signaling system have been
mapped and their biological functions are relatively well understood,
mechanistic information on how WNT signals are molecularly
interpreted by cells and translated into an intracellular response
driving, for example, cell proliferation, migration of differentiation
remains unclear. Since the field of WNT signaling has developed mainly
with an origin in oncology and developmental biology, pharmacological
aspects of this signaling system such as quantification of ligand–
receptor binding and selectivity, molecular details of receptor
activation and activation dynamics, and a quantitative understanding of
ligand–receptor interaction in relationship to the amplitude of
downstream effects are only slowly evolving.
Given the prominent association of WNT signaling with human
disease, the concept of targeting this signaling system
pharmacologically and therapeutically has been pursued intensively
over the years – in part with substantial success defining targetable
components in the involved signaling cascades. However, it must be
underlined that modulating a central signaling system that is absolutely
essential for tissue renewal and homeostasis as most prominently
exemplified by the rapid turnover of the intestinal epithelium, comes
along with large risks for unwanted side effects.
In this Handbook of Experimental Pharmacology, international
experts in the field provide a timely overview of molecular details of
the signaling pathway ranging from WNT proteins themselves
(Willert/Gross), cell surface receptors, such as Frizzleds (FZD; Schulte),
LDL receptor-like proteins (LRPs; Davidson), and receptor tyrosine
kinase-like orphan receptor (RORs; Mellstedt) over intracellular
components such as the enigmatic scaffold protein Dishevelled (Bryja)
to effector systems, such as the transcriptional regulator β-catenin (van
Amerongen). Emphasis is put on the details of molecular mechanisms of
signal transduction in order to elucidate the molecular pharmacology
of cellular communication. Furthermore, the reader is introduced to the
druggability of different branches of the WNT signaling system
including the planar cell polarity pathway (Gao) as well as the
possibilities to screen for WNT system-targeting compounds from
natural resources (Katanaev). While the potential indications for drugs
modulating WNT signaling are numerous, only some important areas
are exemplified here providing insight into the importance of FZD7 as a
therapeutic target in intestinal cancers (Vincan), the use of WNT-
derived peptides for anticancer therapy (Andersson), the opportunities
of targeting lung disease (Gosens) and osteoarthritis (Nalesso) as well
as Alzheimer’s disease (Inestrosa). Last but not least, the chapter by Lai
and Kahn discusses whether any specific WNT targeting therapeutics
would indeed demonstrate clinical efficacy with an acceptable safety
profile.
As pharmacologists, we hope that this collection of chapters
highlighting druggability of the WNT signaling system, accompanying
challenges and risks as well as technical advances to find new drugs
will further stimulate the development of pharmacological concepts
and pharmacological thinking. We are convinced that understanding
the molecular pharmacology of the WNT receptors will go hand in hand
with an intensified quest for effective therapeutics targeting the WNT
signaling system.
References
Schulte G (2010) International Union of Basic and Clinical
Pharmacology. LXXX. The class Frizzled receptors. Pharmacol Rev
62(4):632–667
van Amerongen R (2012) Alternative Wnt pathways and receptors.
Cold Spring Harb Perspect Biol 4(10)
Willert K, Nusse R (2012) Wnt proteins. Cold Spring Harb Perspect
Biol 4(9):a007864
Gunnar Schulte
Pawel Kozielewicz
Stockholm, Sweden
Contents
Part I Basic Pharmacological Principles of the WNT Signaling
System
Controlling Wnt Signaling Specificity and Implications for
Targeting WNTs Pharmacologically
Pooja R. Sonavane and Karl Willert
Extracellular WNTs:Trafficking, Exosomes, and Ligand–Receptor
Interaction
Julia Christina Gross
LRPs in WNT Signalling
Gary Davidson
Targeting the Receptor Tyrosine Kinase ROR1 by Small Molecules
Mohammad Hojjat-Farsangi, Ali Moshfegh, Johan Schultz,
Martin Norin, Thomas Olin, Anders Ö sterborg and Hå kan Mellstedt
Employing Genetically Encoded, Biophysical Sensors to
Understand WNT/Frizzled Interaction and Receptor Complex
Activation
Pawel Kozielewicz, Hannes Schihada and Gunnar Schulte
Can We Pharmacologically Target Dishevelled:The Key Signal
Transducer in the Wnt Pathways?
Miroslav Micka and Vítězslav Bryja
Zooming in on the WNT/CTNNB1 Destruction Complex:Functional
Mechanistic Details with Implications for Therapeutic Targeting
Saskia Madelon Ada de Man and Renée van Amerongen
An Overview of Potential Therapeutic Agents Targeting WNT/PCP
Signaling
Jin Wang, Di Feng and Bo Gao
Mining Natural Compounds to Target WNT Signaling:Land and Sea
Tales
Vladimir L. Katanaev, Artem Blagodatski, Jiabin Xu,
Yuri Khotimchenko and Alexey Koval
Part II Disease-Specific Targeting of the WNT Signaling System
Frizzled7 Activates β-Catenin-Dependent and β-Catenin-
Independent Wnt Signalling Pathways During Developmental
Morphogenesis: Implications for Therapeutic Targeting in
Colorectal Cancer
Bang Manh Tran, Dustin James Flanagan, Toby James Phesse and
Elizabeth Vincan
Targeting Oncogenic WNT Signalling with WNT Signalling-Derived
Peptides
Vikas Yadav, Njainday Jobe, Lubna Mehdawi and Tommy Andersson
WNT Signalling in Lung Physiology and Pathology
Yan Hu, Chiara Ciminieri, Qianjiang Hu, Mareike Lehmann,
Melanie Kö nigshoff and Reinoud Gosens
WNT Signalling in Osteoarthritis and Its Pharmacological
Targeting
Anna De Palma and Giovanna Nalesso
WNT Signaling Is a Key Player in Alzheimer’s Disease
Nibaldo C. Inestrosa, Cheril Tapia-Rojas, Waldo Cerpa,
Pedro Cisternas and Juan M. Zolezzi
Pharmacologically Targeting the WNT/β-Catenin Signaling
Cascade:Avoiding the Sword of Damocles
Keane K. Y. Lai and Michael Kahn
Part I
Basic Pharmacological Principles of the
WNT Signaling System
© The Author(s), under exclusive license to Springer Nature Switzerland AG 2021
G. Schulte, P. Kozielewicz (eds.), Pharmacology of the WNT Signaling System,
Handbook of Experimental Pharmacology 269
https://doi.org/10.1007/164_2021_529
Controlling Wnt Signaling Specificity

and Implications for Targeting WNTs
Pharmacologically
Pooja R. Sonavane1 and Karl Willert1
(1) Department of Cellular and Molecular Medicine, University of
California San Diego, La Jolla, CA, USA
Karl Willert
Email: kwillert@health.ucsd.edu
1 Introduction
2 Wnt Processing and Secretion
3 Wnt Signaling
4 Wnt Signaling Specificity
5 Engineered Specificity
6 Targeting Wnt in Disease
6.1 Cancer
6.2 Tissue Regeneration and Repair
7 Concluding Remarks
References
Abstract
Wnt signaling is critical for proper development of the embryo and for
tissue homeostasis in the adult. Activation of this signaling cascade is
initiated by binding of the secreted Wnts to their receptors. With the
mammalian genome encoding multiple Wnts and Wnt receptors, a
longstanding question in the field has been how Wnt-receptor
specificities are achieved. Emerging from these studies is a picture of
exquisite control over Wnt protein production, secretion, distribution,
and receptor interactions, culminating in activation of downstream
signaling cascades that control a myriad of biological processes. Here
we discuss mechanisms by which Wnt protein activities are tuned and
illustrate how the multiple layers of regulation can be leveraged for
therapeutic interventions in disease.
Keywords Engineered Wnt agonist – Extracellular environment –

Frizzled/Fzd – Lrp5/6 – Ptk7 – Receptor–ligand interactions – Ror1/2 –
Ryk – Signaling specificity – Wnt – Wnt mimetic/surrogate
1 Introduction
Wnt proteins are a family of secreted growth factors with critical roles
in organizing and assembling the body plan of all metazoan species.
The defining feature of all Wnts is a nearly invariant pattern of 22
cysteines across 350–400 amino acids that form disulfide linkages to
yield a globular 2-domain protein of approximately 40 kDa, the
structure of which has been likened to that of a hand with a thumb and
index finger capable of grasping the extracellular domains of their
cognate receptors (Janda et al. 2012).
The diverse biological activities attributed to Wnts across all animal
species can be unified as that of symmetry breaking signals, specifying
the polarity of individual cells within developing tissues to eventually
yield the complex patterns that define individual tissues and the whole
organism. This symmetry breaking property derives from the fact that
Wnts act locally and are rarely found at distances greater than a few cell
diameters from their site of synthesis, a property afforded by a
covalently attached lipid (Willert et al. 2003).
This article will focus on these Wnt proteins and how interactions
with a variety of proteins and extracellular structures regulate their
signaling activity and specificity. Upon engaging cell surface receptor
complexes, Wnts activate intracellular signaling cascades that have
been the subject of many studies and the focus of other review articles.
We will also discuss how this complex network of protein interactions
in the extracellular environment has been leveraged to manipulate Wnt
signaling and develop novel approaches to target and exploit this
pathway for therapeutic applications.
2 Wnt Processing and Secretion

Following transcription, Wnts are co-translationally inserted into the
endoplasmic reticulum (ER) where they encounter Porcupine (Porcn), a
membrane-bound and ER-resident O-acyltransferase (Hofmann 2000;
Zhai et al. 2004), which appends a palmitoleate, a 16-carbon
monounsaturated fatty acid, to a conserved serine residue (Takada et
al. 2006). This O-palmitoleation is shared among all Wnts, with the
notable exception of the distantly related Wnt homolog WntD in
Drosophila (Ching et al. 2008; Chu et al. 2013), and is essential for their
activity. Blocking this lipid attachment, either by mutating Porcn
(Biechele et al. 2011; Kadowaki et al. 1996) or by treating with Porcn
inhibitors (e.g., IWP, C59, LGK974) (Chen et al. 2009; Liu et al. 2013;
Motono et al. 2016), prevents Wnt passage through the secretory
pathway and eventual secretion. Since all Wnts are thought to be lipid-
modified, Porcn mutation or inhibition creates an all-Wnt-mutant
condition.
The lipid moiety plays critical roles in Wnt secretion, distribution
and signaling, as revealed by several elegant cell biological and
structural analyses. First, following palmitoleation by Porcn, Wnt
associates with Wntless/evenness interrupted (Wls/Evi; a.k.a., Gpr177)
(Herr and Basler 2012), a protein with a membrane domain resembling
that of G protein-coupled receptors (GPCRs) that is dedicated to
ushering Wnts through the secretory pathway to the cell surface. In this
complex, the palmitoleate protrudes into the lipid bilayer from a Wnt
hairpin (the thumb), which is nestled in a hydrophobic pocket of the
GPCR-like domain (Nygaard et al. 2021).
Of note, mutations of Wls or Porcn produce nearly identical
phenotypes, and conditional knockout alleles of these genes are
commonly used to establish a requirement for Wnt signaling in any
biological process of interest. In humans, PORCN and WLS mutations
are extremely rare, producing pleiotropic conditions that affect
multiple tissues and organs, consistent with Wnts’ broad range of
actions. The PORCN gene is encoded on the X-chromosome, and
mutation of a single copy leads to a condition called focal dermal
hypoplasia (or Goltz syndrome) in females (Grzeschik et al. 2007; Wang
et al. 2007). In the case of WLS, several partial loss of function
mutations have been identified and shown to lead to pleiotropic multi-
organ defects (Chai et al. in press).
Second, once at the cell surface, multiple mechanisms have been
proposed by which the hydrophobic moiety on Wnt is shielded from the
largely aqueous extracellular milieu (Fig. 1), including association with
lipoprotein complexes (Kaiser et al. 2019; Neumann et al. 2009;
Panakova et al. 2005), exosomes that are formed inside multivesicular
bodies (MVBs) prior to release into the extracellular space (Gross et al.
2012; Korkut et al. 2009; Luga et al. 2012), chaperones, such as SWIM
(Mulligan et al. 2012) and Afamin (Mihara et al. 2016), and direct
transport of Wnts via cytonemes (Hsiung et al. 2005; Stanganello et al.
2015). Highlighting once more the importance of the lipid for Wnt’s
tissue distribution, McGough et al. demonstrated that glypicans (Dlp
and Dally in Drosophila, and human homologs GPC3, 4, 5, and 6) bind
the palmitoleate, thus shielding it from the aqueous environment and
permitting spread of Wnts in the extracellular space, and eventually
handing them over to signaling receptors (McGough et al. 2020).
Fig. 1 Wnt secretion, distribution and signaling. Wnt proteins are synthesized in the
endoplasmic reticulum where an acyl moiety is covalently attached to Wnt by
Porcupine (Porcn). These lipid-modified Wnts are escorted by Wntless (Wls/Evi) to
the cell surface where they are bound by proteoglycans, such as Glypican3/4/5/6.
Membrane-associated Wnts can be transported to neighboring cells via cellular
projections called cytonemes. Additionally, carrier proteins, such as SWIM and AFM,
facilitate transport of secreted Wnts. Wnts can be inactivated enzymatically by
Notum or Tiki, or sequestered by Sfrps, Cer, and WIFs to prevent receptor binding.
Once Wnts reach target cells expressing Wnt receptors (Fzd) and co-receptors
(Lrp5/6, Ror1/2, Ryk, and Ptk7), signal transduction leads to activation of β-
dependent or alternative pathways. Dkk and Sost bind Lrp5/6 and prevent Wnt from
heterodimerizing Lrp5/6 with the Fzd receptors, an event necessary for signal
transduction
The third and final act for Wnt’s lipid moiety is illustrated by the co-
crystal structure of Wnt and the extracellular cysteine rich domain
(CRD) of the Wnt-receptor Frizzled (Fzd) (Janda et al. 2012). Here, the
palmitoleate extending from the thumb lies in a hydrophobic grove of
the CRD, which, together with the index finger of the Wnt hand, serves
to grasp the receptor. Structural analyses have demonstrated a
proclivity of Fzd CRDs to form dimers (DeBruine et al. 2017a; Nile et al.
2017), which can be stabilized by unsaturated fatty acids, indicating
that Fzds at the cell surface may exist in multiple states, as monomers
or as dimers, the equilibrium of which is regulated by receptor density,
as well as by Wnt availability.
3 Wnt Signaling
Many review articles, including in this book volume, have been
dedicated to the mechanisms of Wnt signaling downstream of the
receptors. For brevity-sake – and risking oversimplification – we will
pare down Wnt signaling to two pathways, the Wnt/β-catenin pathway
and the alternative Wnt pathway, commonly referred to as the
canonical and non-canonical Wnt pathway, respectively.
Wnt/β-catenin signaling is initiated when a Wnt protein
heterodimerizes Fzd with a co-receptor encoded by Lrp5/6. Compelling
evidence for Wnt-mediated heterodimerization of Fzd and Lrp5/6
derives from the use of engineered recombinant proteins capable of
simultaneously binding each receptor to activate downstream signaling
(further details on these engineered Wnt-like proteins follow below).
The key downstream effector of this pathway is β-catenin, which
becomes stabilized and subsequently enters the nucleus to activate
expression of target genes. The list of known Wnt target genes is
extensive with many being activated in a tissue and context dependent
manner. Axin2 is considered a universal Wnt target gene (Jho et al.
2002; Lustig et al. 2002), a feature that has been leveraged to trace
developmental lineages of cells that have received an early Wnt signal
(van Amerongen et al. 2012a). In contrast, expression of the Wnt target
gene Sp5, a transcription factor that in part acts to rein in expression of
Wnt target genes, including itself, is restricted to cell populations with
broad developmental potential, such as stem cells (Huggins et al. 2017).
In contrast to the Wnt/β-catenin pathway, alternative Wnt signaling
pathways are thought to be initiated by Wnt binding Fzd and multiple
other co-receptors, in particular the receptor tyrosine kinases (RTKs)
Ror1, Ror2, Ryk, and Ptk7, all of which contain pseudokinase domains
(Sheetz et al. 2020) (Fig. 2). The extent to which Wnts heterodimerize
Fzd and these alternate co-receptors is not as well established as it is
for Fzd-Lrp5/6. However, Wnts can bind FZDs without engaging co-
receptors, as well as act in concert with FZD and these atypical RTKs to
promote a variety of downstream effects, including the establishment
of cell polarity across an epithelium, convergent extension (CE) as seen,
for example, during gastrulation and neural tube closure, and other
complex biological processes that establish form and shape during
development. Of note, in contrast to Wnt/β-catenin signaling, which
regulates gene expression, very few robust transcriptional targets have
been identified downstream of these alternative Wnt signaling events.
Rather, these alternate Wnt pathways may act directly on the proteome,
as illustrated by Wnt5a/Ror regulating the stability of Kif26b (Susman
et al. 2017).
Fig. 2 Wnt receptors, co-receptors, and accessory factors. Wnt proteins bind to a
wide range of cell surface receptors, with Fzd1-10 being the primary receptors and
Lrp5/6, Ror1/2, Ryk, and Ptk being critical co-receptors. Accessory receptors such
as Egfr, Grp124, and Reck facilitate signal transduction in a Wnt-specific manner
Wnt5a is a well-established ligand for Ror1 and 2, and loss of
function mutations in Wnt5a phenocopy Ror1/2 double mutants (Ho et
al. 2012; Nomi et al. 2001). Wnt5a has also been shown to draw Fzd
and Ror2 into a complex to promote downstream signal transduction
involving Dvl polymerization and Rac1 activation (Nishita et al. 2010).
Since the extracellular domains of Fzd and Ror are related, it is
conceivable that Fzd-Ror heterodimer formation is structurally similar
to Fzd-Fzd homodimer formation, however, this has not been confirmed
experimentally. Furthermore, receptor oligomerization can occur
independently of the CRDs through interactions of the transmembrane
domains, as shown for Fzd1 (Kaykas et al. 2004) and FZD6 (Petersen et
al. 2017). Wnt5a can also promote Ror1-Ror2 heterodimerization to
recruit guanine nucleotide exchange factors and activate RhoA and
Rac1 (Yu et al. 2016). Although Wnt5a is a frequent participant in these
alternate Wnt pathways, several other Wnts have been found to signal
through Ror1/2, Ryk, and Ptk7, for example, Wnt11 acting through
Fzd7 and Ryk to affect CE in Xenopus (Kim et al. 2008a), Wnt4
interacting with Ptk7 (also known as Otk) to inhibit Wnt/β-catenin
signaling during embryonic patterning of Drosophila (Peradziryi et al.
2011), Wnt9a genetically interacting with Ror1/2 to regulate skeletal
development in mice (Weissenbock et al. 2019), to name a few.
A pervasive misconception is that individual Wnts can be classified
into distinct classes, those that activate the Wnt/β-catenin pathway and
those that activate the alternative Wnt pathway. However, depending on
cellular context, a specific Wnt can couple to either pathway. For
example, Wnt5a has been associated with alternative Wnt signaling,
interacting with Ror1/2 to promote midgut elongation (Ho et al. 2012;
Yamada et al. 2010). However, in certain contexts, such as ectopic
expression of Fzd4 in mouse cells (Mikels and Nusse 2006) or Fzd5 in
Xenopus embryos (He et al. 1997), Wnt5a activates the Wnt/β-catenin
pathway, illustrating that individual Wnt proteins can stimulate
multiple signaling pathways (van Amerongen et al. 2012b).
In the extracellular space, multiple factors directly or indirectly
restrain Wnt signaling activity. For example, several secreted molecules,
such as secreted frizzled related proteins (Sfrp), Wnt inhibitory factor
(Wif), and Cerberus (Cer), act as receptor decoys and bind Wnts to
prevent their interaction with receptors. Other proteins enzymatically
modify and inactivate Wnts, such as Notum, which cleaves the lipid
moiety (Kakugawa et al. 2015), and Tiki (TRABD2A/B), which cleaves
amino terminal amino acids (Zhang et al. 2012). Inhibitory factors, such
as Dkk and Sost, bind Lrp5/6 (Li et al. 2005; Mao et al. 2001) to prevent
Wnt-mediated receptor heterodimerization (Fig. 1). Several of these
negative regulators of Wnt proteins are direct target genes of the Wnt/
β-catenin pathway, thereby establishing negative feedback loops that
are critical to permit progression of developmental processes.
An especially fascinating class of secreted factors influencing Wnt
signaling activity are the R-spondins (Rspo1,2,3,4) (Kim et al. 2008b).
These proteins serve to stabilize Fzd proteins on the cell surface,
thereby sensitizing cells to the incoming Wnt protein. Initially identified
to be ligands for the widely studied intestinal stem cell marker Lgr5 (as
well as the related Lgr4 and 6 receptors) (Carmon et al. 2011; de Lau et
al. 2011), Rspos also bind the membrane-spanning E3 ubiquitin ligases
Znrf3 and Rnf43, thereby sequestering them and consequently blocking
Fzd ubiquitination and downregulation. Although it is clear from
structural studies that Rspo forms a ternary complex with Lgr5 and
these E3 ligases (Zebisch and Jones 2015), Rspo can act independently
of the Lgrs to inhibit Rnf43 and Znrf3 activity (Szenker-Ravi et al.
2018).
4 Wnt Signaling Specificity

The large number of Wnt genes (19 in mammals) with a nearly equal
number of Wnt receptors (10 Fzds and multiple co-receptors, Fig. 2)
raise the question of how signaling specificity is achieved. In vitro
signaling and binding studies suggest that Wnt–receptor interactions
are highly promiscuous, with a single Wnt interacting with multiple
Fzds, and vice versa. For example, Wnt3a can couple through FZD1, 2, 4, 5,
7, 10 to activate β-catenin signaling, whereas FZD5 can efficiently
transduce Wnt1, 3, 3a, 7a/b, and 8a/b signals (Voloshanenko et al.
2017). Using bio-layer interferometry to measure molecular
interactions indicates that Wnt3a binds the CRDs of FZD4, 5, 7, 8 with
single nano-molar affinity and the CRDs of FZD1, 2 with significantly
lower affinities (Dijksterhuis et al. 2015). Confounding signaling and
binding studies are the lack of purified and biologically active Wnts –
although several recombinant Wnts are commercially available only
Wnt3a and Wnt5a have been characterized thoroughly (Mikels and
Nusse 2006; Willert et al. 2003) – as well as Wnts’ inherently poor
physicochemical properties. Overall, insights gleaned from such in vitro
signaling and binding studies provide limited insights into how Wnt–
receptor interactions are regulated in vivo.
In the case of the Drosophila, the Wnt-1 homolog Wingless (Wg)
interacts with both Fzds encoded in the fly genome, Fz and Dfz2, but
with a ten-fold higher affinity to Dfz2 than with Fz (Rulifson et al. 2000).
Mutation of either fz or dfz2 alone does not produce a segment polarity
phenotype, which would be expected for a receptor essential to Wg
signal transduction. Rather, only upon mutation of both fz and dfz2 is a
segment polarity phenotype exposed (Bhanot et al. 1999), indicating
that either receptor, irrespective of Wg affinity, is capable of functioning
as a bona-fide Wg receptor to establish proper segment polarity.
A recently appreciated mechanism to establish signaling specificity
is through the engagement of accessory factors (Fig. 2). Such accessory
factors by themselves may have only weak binding affinities for Wnts.
However, in context with the primary receptors and co-receptors, these
low affinity interactions contribute to a much higher functional affinity,
or avidity, critical for intracellular signalosome formation and signal
transduction.
An especially illustrious and elegant example of this mode of
establishing Wnt signaling specificity is observed during brain
angiogenesis and blood brain barrier (BBB) maturation. Here,
Wnt7a/7b signaling through Fzd/Lrp5/6 uniquely requires the cell
surface cofactors Reck and Gpr124 (Cho et al. 2017; Posokhova et al.
2015; Ulrich et al. 2016; Vallon et al. 2018; Vanhollebeke et al. 2015;
Zhou and Nathans 2014). A combination of Wnt7a/7b and Fzd4/6
protein engineering, cell signaling studies, and protein binding
experiments defined a specific region of Reck, the CC domain, as well as
two regions of Wnt7a/7b critical for Wnt7/Fzd/Gpr124/Reck signaling
(Cho et al. 2019). Isothermal titration calorimetry and single-molecule
force spectroscopy detected low affinity binding (1.2–7 μM) between
synthetic Wnt7a/7b peptides and the Reck CC domain (Eubelen et al.
2018), illustrating that interactions between a Wnt and an essential
accessory factor, such as Reck, need not be high affinity.
Another example of the engagement of an accessory factor to
establish Wnt-receptor signaling specificity is in the development of
haematopoietic stem cells (HSCs) in zebrafish where Wnt9a acts
through Fzd9b (zebrafish have duplicated Fzd9 genes, named Fzd9a/9b)
to activate Wnt/β-catenin signaling and promote the proliferation of
nascent HSCs as they emerge from the haemogenic endothelium in the
aorta (Grainger et al. 2016, 2019). Morpholino-induced knockdown of
Wnt9a was rescued by overexpression of Wnt9a but not by Wnt9b or
Wnt3a, indicating a highly selective requirement for Wnt9a (Grainger et
al. 2016). Essential to this selectivity is the epidermal growth factor
receptor (Egfr), which interacts with the Wnt9a/Fzd9b/Lrp5/6 receptor
complex in an Egf-independent manner and phosphorylates the
carboxy-terminal tail of Fzd9b to promote signal transduction, possibly
through clathrin-mediated receptor complex internalization (Grainger
et al. 2019). In contrast to the aforementioned detectable, albeit weak,
interaction between Wnt7 and Reck, a direct association between
Wnt9a and Egfr has not yet been established or quantified.
As expected, in both systems, brain angiogenesis and HSC
development, deletion of the Fzd CRD abrogates all binding and
signaling. Interestingly, swapping CRDs to create chimeric Fzds showed
only partial reduction in signaling, indicating that the CRD provides
only part of the specificity. Further of note, the roles of Reck and Egfr in
Wnt signal transduction appear independent of their other functions:
Reck’s action in angiogenesis is fully attributable to Wnt7a/7b signaling
and is independent of its other function as a matrix metalloproteinase
inhibitor, whilst Egfr’s role in mediating Wnt9a-Fzd9b signaling does not
require canonical Egfr ligands. Together, these experiments provide
compelling evidence that signaling specificity is achieved, at least in
part, through the repurposing of other cell surface molecules, such as
Reck and Egfr, that increase the avidity of a Wnt for an Fzd-Lrp5/6
receptor complex.
5 Engineered Specificity
The fact that Wnts activate Wnt/β-catenin signaling through
heterodimerizing Fzd and Lrp5/6 has motivated the engineering of
bispecific binders capable of achieving this same feat. By linking the C-
terminal tail of the Lrp5/6 ligand Dkk1 to a single chain variable
fragment designed from the Fzd-specific antibody Vantictumab, which
binds Fzd1, 2, 5, 7, 8, Janda et al. created the first so-called Wnt surrogate
capable of potently activating Wnt signaling (Janda et al. 2017).
Subsequently, several other groups reported the generation of similar
molecules with varying signaling specificities and all promoting β-
catenin stabilization in a variety of biological settings, including human
embryonic stem cell differentiation, osteogenic lineage commitment of
mesenchymal stem cells, hepatocyte proliferation, intestinal stem cell
proliferation in organoids, restoration of retinal barrier function, and
more (Chen et al. 2020; Chidiac et al. 2021; Dang et al. 2019; Gumber et
al. 2020; Miao et al. 2020; Tao et al. 2019). In an extension to this
approach, coupling the Wnt agonist Rspo to the liver-specific cell
surface protein ASGR1 potently enhanced Wnt signaling and promoted
liver regeneration (Zhang et al. 2020b).
These engineered Wnt-like proteins – variably referred to by the
authors as Wnt surrogates, Fzd and Lrp5/6 agonists (FLAgs), and Wnt
mimetics – exhibit several important advantages over recombinant Wnt
proteins. First, as mentioned above, in cell culture settings native Wnt
proteins interact promiscuously with their receptors, which is partially
overcome in vivo through tissue-specific expression and engagement of
accessory factors, such as Reck and Gpr124 for Wnt7a/7b and Egfr for
Wnt9a. In contrast, these Wnt-like proteins can be engineered to
interact with greater selectivity for cell surface receptors, thus
potentially overcoming the need for specificity-conferring accessory
factors.
A second advantage of these Wnt-like proteins is that they can be
assembled using recombinant proteins with favorable biochemical
properties. Purification of native Wnt proteins has proven extremely
difficult, requiring 3 to 4 chromatography steps with yields of <0.1 mg
per liter of Wnt conditioned medium (Willert 2008). Furthermore, the
lipid-modified Wnt proteins are highly hydrophobic, necessitating the
addition of detergents to maintain their solubility in aqueous
conditions. In contrast, a Wnt mimetic can be entirely comprised of
engineered proteins, such as immunoglobulin sequences or nanobodies
that are soluble in biologically compatible buffers and can be purified
using a single affinity binding step with yields similar to those for
recombinant antibodies.
The use of engineered Wnt-like proteins has provided mechanistic
insights into downstream pathway activation, in particular the role of
receptor oligomerization to promote intracellular signalosome
formation, as previously proposed (DeBruine et al. 2017b). Structural
studies revealed that Wnts can dimerize Fzd CRDs either in a 1:2 or a
2:2 stoichiometry (Hirai et al. 2019; Nile et al. 2017). These higher
order oligomers are hypothesized to promote optimal downstream
signaling, which is supported by observations that bispecific and
monovalent ligands (i.e., capable of binding one Fzd and one Lrp6
molecule) less potently activate downstream signaling than bispecific
and bivalent ligands (i.e., capable of simultaneously binding two Fzd
and two Lrp6 molecules) (Chen et al. 2020; Gumber et al. 2020; Miao et
al. 2020). These findings indicate that higher order receptor
oligomerization is not strictly required for signaling but may control
signal amplitude.
Looking ahead, since all of the aforementioned engineered Wnt
agonists promote Wnt/β-catenin signaling only, it will be of great
interest and value to engineer additional Wnt agonists that target and
activate alternative Wnt signaling pathways.
6 Targeting Wnt in Disease

Given Wnts’ critical roles in embryonic development and tissue
homeostasis, it comes as no surprise that aberrant Wnt signaling is
associated with a host of pathological states. With its many levels of
regulation, both inhibitory and activating, this pathway offers ample
opportunities for therapeutic intervention. Generally speaking, cancer
researchers have sought approaches to inhibit Wnt signaling and target
cancer cells harboring Wnt pathway alterations, with the goal of
arresting malignant cell growth. In contrast, due to Wnts’ critical roles
in stem cell biology, activating this pathway in dormant cells may aid in
regeneration and repair of diseased, defective, and damaged tissues.
Many of the Wnt protein interaction surfaces are regarded as non-
druggable, and approaches to identify small molecules that interfere
with Wnt protein interactions have largely been unsuccessful. Rather,
recombinant proteins, engineered and native, as well as peptides have
garnered the most successes for targeting Wnts in the extracellular
space. Table 1 highlights various approaches and compounds that have
been developed to manipulate extracellular Wnt signaling in a variety
of settings, and below we expand on some of them in the context of
cancer and regeneration.
Table 1 Compounds to target extracellular Wnt signaling
Name Target and MOA Clinical development References

Wnts
Wnt3a (ART352-L) Liposomal Phase1/2 study in (Minear et
formulation of patients undergoing al. 2010)
recombinant spinal fusion
Wnt3a to activate (NCT04378543)
Wnt/β-catenin
signaling
Antibodies
Anti-DKK1 (DKN-01 and Binds and inhibits Phase 1/2 study in (Goldstein et
BHQ880) DKK1 patients with al. 2016;
hepatocellular Iyer et al.
carcinoma and Wnt 2014)
pathway alterations
(NCT03645980);
phase 2 study in
biliary tract cancer
(NCT04057365),
metastatic gastric
cancer
(NCT04166721),
prostate cancer
(NCT03837353), and
advanced solid tumors
(NCT04681248)
Anti-FZD Inhibits Wnt Phase 1 study to (Gurney et
(Vantictumab/OMP- signaling by assess safety, al. 2012)
18R5) binding FZD1, 2, 5, tolerability, and PK in
7, 8 metastatic breast
cancer
(NCT01973309),
NSCLC
(NCT01957007), and
pancreatic cancer
(NCT02005315)
Anti-FZD10 (OTSA101) Antibody alone Phase 1 study in (Fukukawa
has weak patients with relapsed et al. 2008)
antagonistic or refractory synovial
activity, but sarcoma
radiolabeled (NCT04176016)
version showed
significant
antitumor activity
PTK7-ADC (cofetuzumab PTK7-targeted Phase 1 study to (Damelin et
pelidotin/PF-06647020) ADC induced evaluate safety and al. 2017)
sustained tumor efficacy in triple
regressions and negative breast cancer
outperformed (NCT03243331) and
standard-of-care in PTK7-expressing
chemotherapy NSCLC
(NCT04189614)
Anti-ROR1 Blocks Wnt5a Phase 1 study (Choi et al.
(Cirmtuzumab/UC-961) signaling through targeting CLL 2018)
ROR1 (NCT04501939),
metastatic breast
cancer
(NCT02776917)
ROR1-ADC (VLS-101) Selectively kills Phase 1 study (Vaisitti et
cancer cells targeting al. 2021)
overexpressing haematological
ROR1 malignancies
(NCT03833180),
phase 2 for metastatic
solid tumors
(NCT04504916)
Anti-RSPO3 Inhibits Wnt Phase 1, advanced (Salik et al.
(Rosmantuzumab/OMP- signaling by relapsed and 2020)
131R10) binding RSPO3 refractory solid
and thereby tumors and metastatic
blocking its CRC (NCT02482441)
interaction with
LGR4
Anti-SOST Inhibits SOST to FDA approved for use
(Romosozumab, enhance in women with high-
Blosozumab) endogenous Wnt risk osteoporosis.
signaling to Phase 2
promote bone (NCT04232657) and
formation phase 4
(NCT04597931) to
treat SCI
Anti-WLS/GPR177 Blocks Wnt NTD (Seo et al.
secretion and 2018)
tumor growth
Anti-Wnt1 Blocks Wnt1 and NTD (He et al.
induces apoptosis 2004; Rhee
in various cancer et al. 2002)
cells
Anti-Wnt2 Blocks Wnt2 and NTD (You et al.
induces apoptosis 2004)
in melanoma and
NSCLC
Anti-Wnt10b Blocks Wnt10b NTD (Rhee et al.
and decreases 2002)
proliferation of a
HNSCC cell line
Engineered proteins
FZD8CRD-IgG1 Decoy receptor Multiple phase 1 (Fischer et

(Ipafricept/OMP-54F28) that sequesters studies targeting solid al. 2017)
Wnts and tumors
prevents their (NCT02069145,
ability to engage NCT02092363,
receptors NCT02050178,
NCT01608867)
Anti-ROR1 CAR Targeting of Phase 1 study of CAR (Wallstabe
genetically T-cell therapy of et al. 2019)
engineered advanced ROR1+
autologous T- tumors
lymphocytes to (NCT02706392)
ROR1+ tumors
Anti-ASGR1-RSPO2-RA Bi-specific protein NTD (Zhang et al.
that activates Wnt 2020b)
signaling by
targeting RSPO2
to ASGR1-positive
liver cells
Peptides
Wnt5a peptide (Foxy-5) Wnt5a agonist to Phase 2 study in (Säfholm et
block cancer cell subjects with Wnt5a- al. 2006)
migration and low colon cancer
invasion NCT03883802
Wnt5a peptide (Box5) Wnt5a antagonist NTD (Jenei et al.
2009)
FZD7 targeting peptide Blocks Wnt3a NTD (Hansen et
(dFz7-21) signaling and C. al. 2019;
difficile toxin B Nile et al.
(TcdB) binding to 2017)
FZD7
Wnt3a binding peptide Macrocyclic NTD (Otero-
(WAp-D04 [Wnt–AFM- peptide that Ramirez et
peptide-D04]) blocks Wnt3a- al. 2020)
Afamin signaling
DKK1 peptide Blocks DKK1 NTD (Brogi et al.
binding to LRP6 2017)
Small molecules
PORCN inhibitor Inhibits Wnt Multiple phase 1 (Jiang et al.
(CGX1321, ETC-1922159, processing studies targeting solid 2018; Liu et
RXC004, Wnt974, (palmitoleation) tumors al. 2013;
XNW7201) and secretion (NCT03507998, Madan et al.
NCT02521844, 2016)
NCT03447470,
NCT01351103,
NCT03901950)
ADC antibody drug conjugate, CAR chimeric antigen receptor, CLL

chronic lymphocytic leukemia, CRC colorectal cancer, MOA mechanism
of action, NSCLC non-small cell lung cancer, NTD none to date, SCI spinal
cord injury
6.1 Cancer
Since the discovery of the first mammalian Wnt gene, Wnt1, originally
called int-1, in the context of mouse mammary tumors (Nusse and
Varmus 1982) and demonstration of its oncogenic potential
(Tsukamoto et al. 1988), the Wnt field has been inextricably tied to
cancer biology. The majority of cancer-associated mutations in the Wnt
pathway are in intracellular signaling components, such as APC, Axin
and β-catenin, that control the stability of the downstream effector β-
catenin. For example, a comprehensive molecular characterization of
colorectal cancers (CRC) found Wnt pathway alterations in 93% of
cases, with the majority being biallelic inactivation of APC and
activating mutations of β-catenin (Cancer Genome Atlas Network
2012), leading to stabilization of β-catenin and ensuing activation of
downstream target genes, such as c-Myc (He et al. 1998), a regulator of
proliferation and cell cycle progression.
Mutations in Wnt genes are largely loss-of-function and produce
developmental defects rather than promote cancer. Instead, cancer-
associated Wnt pathway alterations upstream of receptors include
overexpression of Wnt genes and silencing of Wnt antagonists,
underscoring opportunities for therapeutic intervention in cancer
through Wnt pathway inhibition. As mentioned above, Wnt processing
and secretion requires the activities of Porcn and Wls, and interfering
with either gene produces all-Wnt mutant phenotypes. Porcn inhibitors
have proven to be powerful allies in the treatment of the so-called Wnt-
addicted cancers. In particular, tumors harboring mutations in the
membrane-bound E3 ubiquitin ligases Rnf43 and Znrf3, which
downregulate Fzd cell surface expression, are highly susceptible to
Porcn inhibitors (Jiang et al. 2013; Koo et al. 2015; van de Wetering et
al. 2015; Yu et al. 2020). Likewise, Porcn inhibitors potently disrupt
growth of Rspo3-translocated cancers (Madan et al. 2016), which have
been observed in 10% of CRCs (Seshagiri et al. 2012). Porcn inhibitors
are also effective in cancers that are driven by Wnt overexpression in
the tumor microenvironment, as observed in the MMTV-Wnt1 mouse
model of mammary tumorigenesis (Proffitt et al. 2013) and in lung
adenocarcinomas where Porcn-positive niche cells promote Wnt
signaling (Tammela et al. 2017).
Aside from bone toxicity (Funck-Brentano et al. 2018), which can be
reduced with alendronate therapy (Madan et al. 2018), Porcn inhibitors
are surprisingly well tolerated, including in the intestine, which
requires continuous Wnt secretion from subepithelial myofibroblasts
(Greicius et al. 2018). The lack of gut toxicity is explained by high
expression levels of drug transporters, such as Mrp1 and Abcc1 that
lower intracellular Porcn inhibitor levels, in the Wnt secreting intestinal
myofibroblasts, a mechanism that presumably evolved to protect the
intestinal stem cell niche from environmental insults (Chee et al. 2018).
Small molecule Wls inhibitors have not yet been developed,
however, one study demonstrated that a monoclonal Wls antibody
could reduce Wnt secretion, perhaps through blocking retrograde
transport of Wls to the ER via the retromer complex, and inhibit
proliferation of gastric cancer cells and suppress tumorigenesis vivo
(Seo et al. 2018). The recent description of a Wls-Wnt co-crystal
structure (Nygaard et al. 2021) will likely aid in the development of
drugs to block Wls function.
Additional approaches to interfere with Wnt proteins is through the
use peptides and proteins, native or engineered, that bind, sequester, or
modify Wnts. For example, several Wnt-specific antibodies have been
developed and shown to block various aspects of cancer cell growth,
mostly in cell culture (Hanaki et al. 2012; He et al. 2004; Rhee et al.
2002; You et al. 2004). However, to date, no Wnt-specific antibodies
have advanced to clinical studies or been approved for clinical use. A
decoy receptor comprised of the FZD8 CRD fused to the Fc portion of a
human immunoglobulin G1 (IgG1), called Ipafricept (OMP-54F28),
binds and effectively sequesters Wnt proteins (Fischer et al. 2017), and
Phase 1 studies in hepatocellular, ovarian, and pancreatic cancers have
been completed, however, significant bone toxicity was observed at
efficacious doses, thus limiting further development (Moore et al.
2019).
Another viable approach is to interfere with Rspo, secreted proteins
that potently augment Wnt activity through upregulation of Fzds. An
anti-RSPO3 antibody (Rosmantuzumab/OMP-131R10) did not progress
beyond Phase 1 studies for treatment of advanced solid tumors,
however, this antibody may be efficacious for other indications, such as
attenuation of liver fibrosis (Zhang et al. 2020a).
Targeting Wnt receptors in cancer has also been explored. A pan-
specific FZD antibody that binds FZD1, 2, 5, 7, 8, called Vantictumab/OMP-
18R5 potently inhibited Wnt signaling and growth of xenografted
FZD7-positive tumors (Gurney et al. 2012). Phase 1 clinical studies
were terminated due to adverse side effects, in particular in bone,
which were likely due to lack of targeting the pathway in an FZD
subtype selective manner, a problem that may be overcome by
developing antibodies with greater selectivity for individual FZDs.
Limited expression in adult tissues or onco-fetal expression profiles,
as is the case for ROR1 (Fukuda et al. 2008), can be exploited with
genetically engineered autologous T-lymphocytes carrying chimeric
antigen receptors (Berger et al. 2015). Wnt receptors with tumor cell-
restricted expression can also be targeted with antibody drug
conjugates, as is the case for ROR1 (Vaisitti et al. 2021) and Ptk7
(Damelin et al. 2017).
Although Wnt pathway activation is generally associated with
tumorigenesis, there are exceptions, as illustrated by Wnt5a and Dkk1.
Loss of Wnt5a, which is linked to alternative Wnt signaling pathways
and capable of antagonizing Wnt/β-catenin signaling, either by
mutation or downregulation, is associated with haematopoietic
malignancies, indicating a tumor suppressive function for Wnt5a (Liang
et al. 2003). Paradoxically, Wnt5a overexpression has also been
observed in multiple cancers, including breast, prostate, skin, and
stomach, where it promotes cell migration and invasion in cancer cell
lines, effects that can be blocked with anti-Wnt5a antibodies (Hanaki et
al. 2012) or with Box 5, a butyloxycarbonyl-modified Wnt5a-derived
hexapeptide (Jenei et al. 2009). Conversely, a related hexapeptide,
called Foxy-5, can mimic Wnt5a signaling and block migration and
invasion of breast cancer cells (Sä fholm et al. 2006). The disparate
effects of Wnt5a in cancer can be attributed in part to differences in the
tumor microenvironment, as well as to distinct Wnt5a isoforms
expressed from alternative promoters to produce Wnt5a proteins with
differing amino-termini (Bauer et al. 2013; Huang et al. 2017).
Dkk1, a well-established Wnt antagonist that binds the co-receptors
Lrp5 and 6, offers another paradoxical example: on the one hand, Dkk1
acts as a tumor suppressor by inhibiting Wnt/β-catenin signaling and,
on the other hand, it can promote tumor growth. One mechanism to
explain Dkk1’s tumor promoting activity is through its interaction with
cytoskeleton-associated protein 4 (Ckap4), leading to AKT activation
and promoting cancer cell proliferation (Shinno et al. 2018).
Furthermore, Dkk1 has well-documented immunomodulatory
properties (D'Amico et al. 2016; Malladi et al. 2016). These
observations have spurred the development of neutralizing DKK1
antibodies, including DKN-01 and BHQ880, both of which entered
clinical studies for the treatment of a variety of malignancies.
6.2 Tissue Regeneration and Repair

Wnts’ roles in regeneration and repair have been documented in a
variety of model organisms across the animal kingdom. With several
emerging technologies, it is now possible to harness this regenerative
potential in clinical settings. As mentioned above, Wnt proteins exhibit
poor physiochemical properties, which have complicated the
development of Wnt formulations suitable for clinical application. One
promising approach has been to incorporate Wnts into liposomal
complexes and thereby shield Wnt’s lipid moiety in an otherwise
aqueous environment. This approach increased Wnt3a’s signaling
duration and strength compared to unassociated and detergent
solubilized Wnt3a (Morrell et al. 2008; Tuysuz et al. 2017). Importantly,
liposomal Wnt3a could be delivered in vivo where it promoted bone
growth (Minear et al. 2010) and increased bone formation around bone
implants (Coyac et al. 2020; Li et al. 2021), findings that have led to
clinical studies in which patients receive a liposomal Wnt3a application
following spinal fusion surgery.
A limitation of these liposomal formulations is that they require
initial purification of native Wnt proteins, and the majority of Wnts has
eluded biochemical isolation in biologically active form. Here, the
aforementioned Wnt mimetics/surrogates provide new opportunities
to selectively and specifically activate Wnt-receptor complexes. In
addition, tissue targeted Rspo mimetics, as demonstrated for the
bispecific anti-ASGR1-RSPO2-RA molecule in liver regeneration (Zhang
et al. 2020b), will likely soon advance to clinical studies.
With Wnt’s critical roles in bone homeostasis, Wnt augmentation in
degenerative bone disorders, such as osteoporosis, has proven
successful. Deficiency of the secreted protein Sclerostin (SOST), a Wnt
antagonist that binds Lrp5/6 (Li et al. 2005), leads to sclerosteosis, a
condition characterized by markedly thickened bones (Balemans et al.
2001). Treatment with SOST blocking antibodies, including
Romosozumab and Blosozumab, increased bone mass in a variety of
preclinical animal models of osteoporosis (Ominsky et al. 2017), and in
2019, the FDA approved Romosozumab for treatment of osteoporosis
in postmenopausal women with high risk of bone fractures.
Interestingly, SOST antibody treatment can lead to DKK1 upregulation
through a negative feedback loop, thus limiting Wnt-promoted bone
formation. To overcome this effect, a bispecific antibody to
simultaneously neutralize SOST and DKK1 led to a synergistic effect on
bone formation (Florio et al. 2016). Such drugs will likely offer new
opportunities in combating degenerative bone disorders.
With Wnts’ well-documented roles in a variety of pathologies, in
particular cancer, therapeutic application of Wnt activators to promote
tissue regeneration and repair is potentially problematic. Ex vivo
manipulation of cells and tissues with Wnt activators to promote their
expansion and differentiation prior to replacement therapy will
sidestep such risks.
7 Concluding Remarks
The nearly four decades of extensive research in the Wnt field have
yielded important insights into a broad spectrum of biology and across
the entire animal kingdom. With their role as symmetry breaking
signals, Wnts and their downstream signaling cascades participate in
the establishment and organization of cell and spatial identity. The
complex layers of inhibitory and stimulatory regulation that fine-tune
Wnt activities offer fertile ground for continued scientific investigation
and discovery, as well as countless opportunities for clinical
development of therapies that target cancer and degenerative
disorders. To avoid adverse side effects in the clinic, such as bone and
gut toxicity, it will be critical to devise approaches that activate or
inhibit Wnt pathways with great precision, selectivity and specificity.
Recent technological advances in biomedical research in general and in
the Wnt field specifically are paving the way towards new therapies for
currently incurable diseases.
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Another random document with
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migrations; in most cases they are undertaken simply to gratify a
passing desire.
Quite otherwise is it with those mammals which, every year about
the same time, leave their habitat for some other region often far
distant, from which at a definite time they will return to their former
abode. These migrate; for they do not seize a chance opportunity,
but obey, consciously or unconsciously, a compelling necessity.
The fundamental cause of all true migration among mammals, is
some very distinct and decided seasonal change. In countries of
everlasting spring true migrations do not take place, for want is never
imminent. Summer must contrast with winter, whether the latter bring
frost and snow, or heat and drought; scarcity must alternate with
superfluity before the sluggish mammal makes up its mind to
migrate.
To a slight extent migrations take place among all mountain animals.
The chamois, the steinbock, the Alpine hare, the marmot all migrate
when the snow begins to melt, or a little later; they clamber over
hillsides and glaciers to the heights above, where the pasturage,
now laid bare, promises rich and abundant nourishment, and they
return to the lower slopes of the mountain before winter sets in. The
bear, by nature omnivorous, by habit a thief, undertakes a similar
migration at the same season, and completes it before winter sets in,
—at least so it is in the mountains of Siberia; the various wild cats
and dogs which live among the mountains do the same. Such
changes of residence occur also on the mountains of southern
countries, even of those lying within the tropics. In India and Africa
certain species of monkey ascend and descend the mountains at
regular intervals; elephants seek the high grounds on the approach
of summer, the low grounds in winter; on the Andes in South
America the guanacos flee before the snow into the valleys, and
before the summer-heat to the shoulders of the mountains. All these
migrations are confined by the mountains within comparatively
narrow limits. They only involve a change of altitude of from three to
nine thousand feet, or a journey which may be accomplished in a
few hours, or, at most, in a few days. They have, however, the
regularity characteristic of true migrations, especially in the precise
periodicity of their occurrence, and not less in the constant choice of
the same routes.
Highlands and plain, sea and air, offer a much wider field than the
mountains, and therefore the migrations of the animals inhabiting, or
temporarily traversing these can be more easily observed, and they
are more appropriately termed migratory animals than the dwellers
among the mountains. In the tundras of Russia and Siberia, the
reindeer, which, in Scandinavia, never leaves the mountains,
migrates to a great distance every autumn and returns the following
spring to his former summer haunts.[57] About the same time it
leaves Greenland, and, crossing the sea on a bridge of ice, reaches
the continent of America, where it spends the whole winter, only
returning to the hills of its native peninsula the following April. In both
cases, dread of the approaching winter does not seem to be the sole
cause of migration; there is at the same time a further incentive
supplied by a plague much feared in the far north. For the short
summer on these expanses calls to life an insect-world poor in
species, but endlessly rich in individuals, particularly an
indescribable number of mosquitoes and bot-flies, which make life a
burden to the reindeer, as well as to man. To escape these the
reindeer forsakes the marshy tundra, over which dense clouds of
mosquitoes hover during summer, and hies to where the scourge is
less severely felt—to the Alpine heights, which, in the summer
season afford their most fragrant pasturage. From inherited habit,
the reindeer migrate not only at the same time, but along the same
paths, thus forming tracks which may be distinctly traced, traversing
the tundra for many miles, and crossing streams and rivers at
definite places. At the beginning of the journey, the cows with their
calves arrange themselves in herds of from ten to a hundred, and
precede the young stags and hinds, which are followed again by the
old stags. One troop follows directly behind another, and the
observer can count thousands as they pass. All hurry incessantly on,
turning aside neither for the mountains nor the broad streams which
cross their path, and resting only when they have reached their
winter-quarters. Packs of wolves, bears, and gluttons follow close on
their heels and often pursue them no small part of the way. In spring,
on the return journey, the animals keep to the same order, but the
herds are much smaller, and they travel in a much more leisurely
fashion, and keep less strictly to the paths by which they went.
Journeys still longer than those of the reindeer are taken by the
American bison, the “buffalo” of the prairies.[58] What distance
individual animals travel cannot be stated with certainty, but herds in
course of migrating have been met from Canada to Mexico, from the
Missouri to the Rocky Mountains, and it may be assumed that a
single herd traverses a considerable part of the country lying
between these limits. The bisons have been seen in summer
scattered over the boundless prairie, and in winter in the same
places, but assembled in many thousands; their migrations have
been observed, for they have been followed for hundreds of miles
along the tracks—the so-called “buffalo-paths”, trodden out straight
across plains and over mountains. We learn from eye-witnesses that
a stream a mile wide is to them no barrier, scarcely even a
hindrance, for they throw themselves into it like an irresistible
avalanche, so that the water is covered with the dark, moving throng;
that the animals associate and separate again, the herds increase
and diminish; that old, surly, tyrannous, malevolent bulls avoid the
other bisons, having perhaps been expelled from the herd, and
compelled, probably only after protracted struggles, to live in hermit-
fashion until the following summer; and that, during heavy snowfalls,
the herds take shelter in the forests or on the slopes of the
mountains. From July onwards they begin their migrations from the
north towards the south. Small companies, which, till then, have
been leading a comfortable summer life, combine with others and set
out on the journey with them; other troops join the band, which grows
as it presses on, until there is, at length, formed one of those
extraordinary herds which, united till the next spring, moves and acts
as if animated by one soul. When the winter is safely past, the army
gradually breaks up, probably in exactly reverse order, into herds,
and these divide more and more until at length only small companies
are left. This breaking-up takes place during the course of the return
journey. Both in going and returning, one herd follows another at
some distance, but more or less along the same paths. Specially
favourable places, such as low grounds covered with rich grass,
cause a temporary damming up of the living stream. In such places
incalculable herds assemble together, spend days in the same spot,
and break up only again when all the grass has been eaten, and
hunger urges them to continue their journey. As they march the
wolves and bears follow their track, while eagles and vultures, birds
of ill omen, circle over their heads.
Fig. 36.—A Herd of American Bison or Buffalo.
Scarcity of water, as well as of food, is often a cause of regular

migrations. When winter approaches in the south-east of Siberia,
more particularly in the high Gobi steppe, all the non-hibernating
mammals are compelled, by the peculiar circumstances of these
highlands, to seek refuge in lower-lying regions. The winter in these
high grounds of Central Asia is not more severe than in districts lying
further to the north or north-east, but it is usually almost snowless,
and such pools of water as have been formed by the extremely slight
fall of rain or snow, are covered with a thick sheet of ice. As soon as
this sheet becomes so strong that the animals inhabiting the Gobi
are unable to break it, they are obliged to change their quarters, and
they travel not only to southern but to northern lands, whose only
advantage is that they are covered with snow, for this affords ready
refreshment to the parched tongues of the wanderers, and offers
less resistance to their weak feet than the hard, unbreakable, and
less easily melted ice. This is the explanation of the fact that the
antelope, of which great numbers are found in the Gobi, forsakes a
land which, save for the lack of snow and therefore of available
water, is exactly the same as that which it chooses for its winter
quarters. Not hunger, but thirst, drives it from its home. At the
beginning of winter, the antelopes, at all times gregarious, assemble
in herds of many thousands, spreading over all the low grounds
around their native plateau; they often travel at the rate of fifty or
sixty miles in a single night, and extend their wanderings many
hundreds of miles beyond the boundaries of their proper habitat. The
observer who follows them can detect their tracks everywhere, and
in such numbers that it seems as though vast herds of sheep, far
exceeding in number any ordinary flock, had just passed by.
Fig. 37.—Wild Horses crossing a River during a Storm.
Before the Chinese antelope begins its migration, restlessness

seizes the kulan or dziggetai, probably the ancestor of our horse,
and certainly the most beautiful and the proudest of all wild horses.
The foals of the summer are by autumn strong enough to be able to
endure a long journey with quick marches, and to bid defiance to all
the accidents and dangers of a wandering life. The young stallions
attain their full strength at the end of their fourth year, and towards
the end of September they leave the parent-herd and press forward.
Finally, the impulse to mate begins to animate the older stallions and
mares, and with it comes unrest and the desire to wander. Thus the
fleet, enterprising animals begin their annual migration long before
winter has set in, before even its approach has become at all
apparent; and on this account their migrations at first lack steadiness
and regularity, and have something of the character of journeys in
search of adventure. With the intention of shaking off the
burdensome yoke of the leader and absolute lord of the herd, and of
becoming independent and in their turn equally despotic, the young
stallions forsake the herd, and thenceforward traverse the sandy
steppes singly. All the younger mares who are mature, and even
many of the older ones, seem to be animated by the same feeling as
the young stallions, and they attempt to escape from the rule of their
tyrant and join his young rival, to fall immediately under his dominion.
But not without a struggle does the new candidate for leadership
gain his troop of mares; the old leader does not readily relinquish his
rights. For hours together the stallion stands on the top of a hill or on
the shoulder of a ridge, keenly scanning the country around. His eye
wanders over the desert, his dilated nostrils are turned towards the
wind, his ears are directed forwards on the alert. Eager for battle, he
rushes at full gallop towards every herd which approaches, every
adversary who shows himself; and a furious struggle takes place for
the possession of the mares, who always attach themselves to the
victor. Such combats and strife set the herd in motion, detach it from
the place where the summer has been spent, and lead on to
migrations which become gradually regular, prolonged, persistent,
and almost uninterrupted. In the course of these, if not before the
end of the combats just described, the kulan troops assemble in
ever-growing numbers, until at length herds of more than a thousand
head set out together for fields which give promise of pasturage.
They do not break up while in their winter quarters, and they are thus
compelled to be continually on the move in order to find sufficient
nourishment. The combined tread of the army, as they gallop on in
their usual furious fashion, rings dully out, and more than once, in
Russia, the sound has called the Cossacks of the military cordon to
arms. No wolf ventures to attack such a herd, for the courageous
wild horses know so well how to use their hoofs against him that he
soon gives up any attempt; it is only the sick and exhausted horses
which become his prey, as he follows the wandering herd. Even man
can do them no great damage, for their caution and shyness render
them difficult of approach. But winter, especially if much snow falls,
brings them much suffering. The pasture, at all times scanty, is
exhausted the more quickly the more numerous the herd which
feeds on it. Then the animals devour indiscriminately all the
vegetable substances they can find. For months together they have
to maintain life on leafless shoots. Their bodies cease to be fat and
plump, till at length they are like wandering skeletons. The mother,
herself starving, is no longer able to nourish her foal, for the milk-
yielding udder dries up in times of such need. Many a one whose
tender youth is unable to endure the hard fare dies of starvation.
Even the old horses suffer from the poverty and treachery of the
winter. Snow-storms blowing over their feeding-ground for days at a
time depress their usually cheerful courage, and increase the
boldness of the wolves, which, even if they do not fall upon the
already exhausted horses, persecute and annoy to the utmost those
who are not yet worn out. But as soon as circumstances begin to
improve, the wiry, weather-hardened, enduring creatures recover
their high spirits, and, when the snow begins to melt, they set out on
the return journey, reaching their summer home in about a month’s
time. There they break up into single herds, recuperate among the
luxuriantly sprouting, fragrant pasture, and, in a surprisingly short
time, become fat and plump again. Soon the want and misery of the
winter are forgotten.
Great as are the distances often traversed by all the mammals
already mentioned, they can scarcely be compared with those
covered by seals and whales. The water favours all the movements
of animals adapted to aquatic life, and offers everywhere the same
general conditions of life and the same amenities. Thus it renders
the migrations of its inhabitants easier, less toilsome and hazardous
than those of any other wanderers. Nevertheless it is somewhat
surprising to learn that many sea-mammals, and particularly the
whales, are among the most nomadic of all creatures; in fact that
many, if not most of them, pass their whole life in travelling. Strictly
speaking, no whale has a permanent place of sojourn for the whole
year, but passes singly, in pairs, with its young, or in more or less
numerous companies—the so-called schools—from one part of the
ocean to another, visiting certain favourite haunts in regular order,
and choosing different haunts in summer and in winter. The seas
inhabited by the same species of whale in winter and in summer
often lie farther apart than people seem to suppose, for some whales
travel, twice a year, more than a quarter of the earth’s circumference;
they are to be found in summer among the ice-floes of the Arctic
Ocean, and in winter on the other side of the Equator. The female
whales, who are in the highest degree sociable, and attached to their
young with the tenderest, most devoted love, assemble together in
surprising numbers, and under the guidance of a few males, traverse
the ocean by definite routes and at definite times, some keeping to
the open sea, others making their way along the coasts. Storms may
force them to change their route, or delay in the appearance of the
animals on which they feed, whose occurrence and disappearance is
obviously the chief cause of their migration, may to some extent
influence their course and the time of their visiting certain spots; but,
as a general rule, their migrations are so systematic that on northern
and southern coasts people look for the arrival of the whale on a
particular day, and place watches so that they may be able to begin
the long-desired chase without loss of time. Whales, which are
recognized by the dwellers on the coast by some mark, such as
mutilated fins, and which have been several times pursued in vain,
have been known to appear several years in succession at the same
time and at the same place; and the chase after these most valuable
and therefore increasingly persecuted animals takes place with the
same regularity as do hare-hunts on land, though at any other time
of the year it would be vain to look for them. “After Twelfth Day,” says
old Pontoppidan, “the Norwegians watch from all the hills for the
whale, whose arrival is announced by the herring.” First appears the
killer, then, three or four days, or at the most a fortnight later, the
rorqual, though, apparently, one comes from Davis Straits and the
other from Greenland. On the south coasts of the Faroë Islands, and
especially in the Qualbenfjord, from three to six bottle-nose whales
still appear every year about Michaelmas, as they did a hundred and
ninety years ago. In a Scottish bay there appeared twenty years in
succession a rorqual, which was generally known by the name of
“Hollie Pyke”, and was pursued every year and finally captured. On
the coast of Iceland single whales choose the same bays for a
temporary sojourn every year in the same months, and even weeks,
so that the inhabitants have got to know them individually, and have
given them special names. Certain well-known mother-whales visit
the same bays every year to bring forth their young, and they
themselves are spared, but they have to purchase their own lives,
dearly enough, at the cost of that of their young ones, which are
regularly taken captive. It is very unusual for the migrating whales to
keep neither to time nor to route; in general, their journeys are as
regular as if they were arranged according to the position of the
stars, and as if they took place along laid-out paths bounded on both
sides. No other mammal migrates more regularly; indeed, their
wanderings may be compared with the migrations of birds.
The seals, like the whales, migrate every year, on the whole with
great regularity, though not to such a distance. Those species which
inhabit inland seas cannot, of course, leave these, but they traverse
them every year in regular order, or at certain times ascend the
rivers flowing into them; all the ocean species, on the other hand, set
out every autumn and spring, by definite routes, to certain regions or
localities. All the seals in the far north, as well as those in the seas
about the South Pole, are forced to migrate by the extension of the
ice in winter, and may travel with it towards temperate zones,
returning towards the poles again as the ice melts. But they, like all
other members of their order, are impelled to travel for another not
less weighty reason; they require the mainland, or at least large,
spreading, fixed masses of ice, on which to bring forth and nurture
their young, until these are able to follow them into the water, there
to shift for themselves. Thus every year thousands and hundreds of
thousands of seals appear on certain islands and ice-banks,
covering some of these birth-places of their race in such crowds that
every available spot must be utilized in order to secure space for all
to bring forth their young. They pass weeks, even months, on land or
on the ice without hunting, descending into the sea, or taking food;
they suckle their young, then mate, and by degrees break up their
great assemblage, distributing themselves over the wide ocean to
resume their former manner of life, or setting out with their young,
who still require training, on more or less extensive foraging
expeditions.[59]
Fig. 38.—Flying Foxes.

As is well known, there are many mammals which have the habit of
hibernating, which pass the severe part of the year well protected in
deep and carefully closed burrows, and are thus spared the
necessity of leaving their haunts. Even among these, however, at
least among those living in the temperate zones, there are some
which migrate during their waking time, namely, the bats. Defective
as the wing of a bat must appear when compared with that of a bird,
it is nevertheless of such assistance in flight, that it makes journeys
possible which seem out of all proportion to the size of the animal.
Another fact makes travelling easier to the restless bat; it is not tied
down by its offspring to any particular spot, for the young one
attaches itself directly after birth to the breast of the mother, and is
borne by her through the air till it is capable of independent life. The
bat is thus one of the best-adapted of migratory mammals, and,
under some circumstances, it makes full use of its advantages. As a
general rule, the wanderings of the different species of bat are to be
regarded simply as excursions made with a view to taking advantage
of any district which is, for the time being, particularly rich in food; but
they do sometimes become really long journeys, which lead some
species to far distant lands, and they are then not without the
regularity characteristic of all true migrations. The largest bats, the
flying foxes, fly long distances every evening in search of the fruits
on which they chiefly subsist; they do not hesitate to cross an arm of
the sea fifty or sixty miles in breadth, and they must even have
traversed the distance between Southern Asia and the East of
Africa, as certain species occur in both these regions. The bats
proper accomplish at least as much. Following the reappearance of
the insects, which occurs at different times in regions of different
altitude, they ascend from the plains to the mountain heights, and
descend in autumn to the low grounds again; they pursue the
numerous flies which congregate about the wandering cattle-herds
of Central Africa, and they migrate also from the south towards the
north and return southwards again, or in reverse order. The boreal
bat appears at the beginning of the bright nights in the north of
Scandinavia and Russia, and leaves these districts, which may be
considered its head-quarters, towards the end of summer, to spend
the winter among the mountains of Central Germany and the Alps.
The pond-bat is regularly seen on the plains of North Germany
during summer, but only exceptionally at that period among the
mountains of Central Germany, in whose caverns it spends the
winter. That other species of bat occurring in Germany change their
place of abode in a similar manner can scarcely be doubted.
In the cases cited, which have been selected from a mass of
available material, I have given examples of those migrations of
mammals which we may call voluntary, because of their regularity;
but in so doing I have by no means completed my task. Hunger and
thirst, the poverty and temporary inhospitableness of a particular
region, sometimes press so severely on certain mammals that they
endeavour, as if despairing, to save themselves by flight. Abundant
nourishment and good weather favour the increase of all animals,
and affect that of a few plant-eating mammals to such an
extraordinary degree, that, even under propitious conditions, their
habitat must be extended. But if one or more rich years—in some
cases a few favourable months—be followed by a sudden reverse,
the famine soon passes all bounds, and robs the creatures not only
of the possibility of subsistence, but also of all hope, or at least of all
presence of mind.
It is under circumstances such as these that the field-voles of our
own country, and the Siberian voles, assemble in enormous
multitudes, leave their native haunts and migrate to other districts,
turning back for no obstacles, avoiding the water as little as the
forbidding mountains or the gloomy forest, fighting to the last against
hunger and misery, but perishing hopelessly from diseases and
epidemics which rage among them like plagues, reducing armies of
millions to a few hundreds. Thus, too, the squirrels of Siberia, which,
in ordinary years, undertake, at the most, only short excursions,
assemble in vast armies, hurry in troops or companies from tree to
tree, in compact masses from forest to forest, swim across rivers and
streams, throng into towns and villages, lose their lives by
thousands; but suffer no obstacle or hindrance—not even the most
obvious dangers—to delay them or divert them from their path. The
soles of their feet become worn and cracked, their nails ground
down, the hairs of their usually smooth fur rough and matted.
Through the forest lynxes and sables, in the open fields gluttons,
foxes and wolves, eagles, falcons, owls and ravens follow them
closely; pestilence claims more victims from their ranks than the
teeth and claws of beast of prey or the guns and cudgels of men, yet
they press on and on, apparently without hope of return. A Siberian
sportsman of my acquaintance gave me a verbal account of the
appearance of such an army of squirrels, in August 1869, in the town
of Tapilsk, among the Ural Mountains. It was only one wing of a
migrating army, of which the main body travelled through the forest
about five miles farther north. Sometimes in single file, sometimes in
companies of varying strength, but in unbroken succession, the
animals pressed on, crowding as densely through the town as
through the neighbouring forest; used the streets, as well as the
hedges, and the roofs of buildings as paths; filled every court-yard,
thronged through windows and doors into the houses, and created
quite an uproar among the inhabitants—much more among the
dogs, which killed thousands of them, evincing an unbridled
bloodthirstiness till then unsuspected. The squirrels, however, did not
seem to concern themselves in the least about the innumerable
victims falling in their midst; in fact, they took no notice of anything,
and allowed nothing to divert them from their route. The procession
lasted for three whole days, from early morning till late in the
evening, and only after nightfall each day was there a break in the
continuity of the stream. All travelled in exactly the same direction,
from south to north, and those that came last took the same paths as
their predecessors. The rushing Tchussoveia proved no obstacle, for
all that reached the bank of that rapid mountain-river plunged without
hesitation into its whirling and seething waters, and swam, deeply
sunk and with their tails laid across their backs, to the opposite bank.
My informant, who had been watching the procession with growing
attention and sympathy, rowed out into the midst of the throng. The
tired swimmers, to whom he stretched out an oar, climbed up by it
into the boat, where, apparently exhausted, they sat quietly and
confidingly, until it came alongside a larger vessel, when they
climbed into that, and remained on it for some time as indifferent as
before. As soon as the boat touched the bank they sprang ashore,
and proceeded on their journey as unconcernedly as if it had
suffered no interruption.
It must be similar circumstances which compel the lemmings to the
migrations which have been known for centuries. For many
successive years the heights in the tundras of Scandinavia, North
Russia, and the North of Siberia afford them comfortable quarters
and abundant nourishment; for the broad ridges of the fjelds and the
extensive plains between them, the highlands and the low grounds,
offer room and maintenance for millions of them. But not every year
do they enjoy the accustomed abundance for the whole summer. If a
winter in which much snow falls, and which is therefore favourable to
them, as they live safely below the snow, be followed by an early,
warm, and agreeable spring, their extraordinary fertility and power of
increase seem to have almost no bounds, and the tundra literally
teems with lemmings. A fine warm summer increases their numbers
past computing, but it also accelerates the life-course of all the
plants on which they feed, and before it is over these are partly
withered, partly devoured by the greedy teeth of the insatiable
rodents. Scarcity of food begins to be felt, and their comfortable life
comes to an end in panic. Their fearless, bold demeanour gives
place to a general uneasiness, and soon a mad anxiety for the future
takes possession of them. Then they assemble together and begin
to migrate. The same impulse animates many simultaneously, and
from them it spreads to others; the swarms become armies; they
arrange themselves in ranks, and a living stream flows like running
water from the heights to the low grounds. All hurry onwards in a
definite direction, but this often changes according to locality and
circumstances. Gradually long trains are formed in which lemming
follows lemming so closely that the head of one seems to rest on the
back of the one in front of it; and the continuous tread of the light,
little creatures hollows out paths deep enough to be visible from a
long distance in the mossy carpet of the tundra. The longer the
march lasts, the greater becomes the haste of the wandering
lemmings. Eagerly they fall upon the plants on and about their path
and devour whatever is edible; but their numbers impoverish even a
fresh district within a few hours, and though a few in front may pick
up a little food, nothing is left for those behind; the hunger increases
every minute, and the speed of the march quickens in proportion;
every obstacle seems surmountable, every danger trifling, and
thousands rush on to death. If men come in their way they run
between their legs; they face ravens and other powerful birds of prey
defiantly; they gnaw through hay-stacks, climb over mountains and
rocks, swim across rivers, and even across broad lakes, arms of the
sea, and fjords. A hostile company, like that behind the migrating
squirrels, follows in their wake: wolves and foxes, gluttons, martens
and weasels, the ravenous dogs of the Lapps and Samoyedes,
eagles, buzzards, and snowy owls, ravens and hooded crows fatten
on the innumerable victims which they seize without trouble from the
moving army; gulls and fishes feast on those which cross the water.
Diseases and epidemics, too, are not awanting, and probably
destroy more than all their enemies together. Thousands of
carcasses lie rotting on the wayside, thousands are carried away by
the waves; whether indeed any are left, and whether these return
later to their native Alpine heights, or whether all, without exception,
perish in the course of their journey, no one can say with certainty;
but so much I know, that I have traversed great tracts of the tundra of
Lapland where the paths and other traces of a great migrating army
were to be seen almost everywhere, while not a single lemming
could be discovered. Such tracts, I have been told, remain thus for
several successive years, and only after long periods become
gradually repeopled with the busy little rodents.[60]
What hunger causes in the North is brought about by the tortures of
thirst in the richer South. As the brackish pools which have afforded
water to the zebras, quaggas, antelopes, buffaloes, ostriches, and
other animals of the steppes, dry up more and more under the
burning heat of a South African winter, all the animals whose
necessities have hitherto been supplied by the steppes assemble
about the pools which still contain a little water, and these become
scenes of stirring, active life. But when these, too, evaporate, the
animals which have congregated around them are compelled to
migrate, and it may happen that despair takes possession of them,
as it did of the little rodents already described, and that, collecting in
herds like the wild horses and Chinese antelopes (dzieren) of the
steppes of Central Asia, or the bisons of the North American prairies,
they rush straight on for hundreds of miles, to escape the hardships
of winter.
In the South, too, the wild horses are the first to turn their backs on
the inhospitable country. Till the drought sets in, these beautifully-
marked, strong, swift, self-confident children of the Karroo, the
zebra, quagga, and dauw, wander careless and free through their
vast domain, each herd going its own way under the guidance of an
old, experienced, and battle-tried stallion. Then the cares of the
winter season begin to make themselves felt. One water-pool after
another disappears, and the herds which gather about those which
remain become more and more numerous. The general distress
makes even the combative stallions forget to quarrel and fight.
Instead of small companies, herds of more than a hundred head are
formed, and these move and act collectively, and finally forsake the
wintry region altogether before want has enfeebled their powers or
broken their stubborn wills. Travellers describe with enthusiasm the
spectacle presented by such a herd of wild horses on the march. Far
into the distance stretches the sandy plain, its shimmering red
ground-colour interrupted here and there by patches of sunburnt
grass, its scanty shade supplied by a few feathery-leaved mimosas,
and, as far away as the eye can reach, the horizon is bounded by
the sharp lines of mountains quivering in a bluish haze. In the midst
of this landscape appears a cloud of dust which, disturbed by no
breath of air, ascends to the blue heavens like a pillar of smoke.
Nearer and nearer the cloud approaches, until at length the eye can
distinguish living creatures moving within it. Soon the brightly-
coloured and strangely-marked animals present themselves clearly
to the spectator’s gaze; in densely thronged ranks, with heads and
tails raised, neck and neck with the quaintly-shaped gnus and
ostriches which have joined their company, they rush by on their way
to a new, and possibly far-distant feeding-ground, and ere the
onlooker has recovered himself, the wild army has passed by and is
lost from view in the immeasurable steppe.
Fig. 39.—Springbok Antelopes.
The antelopes, which are also driven out by winter, do not always
follow the same paths, but usually travel in the same direction. None
is more numerous or more frequently seen than the springbok, one
of the most graceful and beautiful gazelles with which we are
acquainted. Its unusual beauty and agility strike everyone who sees
it in its wild state, now walking with elastic step, now standing still to
feed, now springing about in playful leaps, and thus disclosing its
greatest ornament, a mane-like snow-white tuft of hair, which at a
quieter pace is hidden in a longitudinal groove of the back. None of
the other antelopes, when forced to migrate, assemble in such
numerous herds as this one. Even the most vivid description cannot
convey to one who has not seen a herd of springboks on their
journey any adequate idea of the wonderful spectacle. After having
congregated for weeks, perhaps waiting for the first shower of rain,
the springboks at last resolve to migrate. Hundreds of the species
join other hundreds, thousands other thousands, and the more
threatening the scarcity, the more torturing the thirst, the longer the
distances which they cover; the flocks become herds, the herds
armies, and these resemble the swarms of locusts which darken the
sun. In the plains they cover square miles; in the passes between
the mountains they throng together in a compact mass which no
other creature can resist; over the low grounds they pour, like a
stream which has overflowed its banks and carries all before it.
Bewildering, intoxicating, and stupefying even the calmest of men,
the throng surges past for hours, perhaps days together.[61] Like the
greedy locusts, the famishing animals fall upon grass and leaves,
grain, and other fruits of the field; where they have passed, not a
blade is left. The man who comes in contact with them is at once
thrown to the ground, and so sorely wounded by the tread of their
hoofs, light indeed, but a thousand times repeated, that he may be
glad if he escapes with his life; a herd of sheep feeding in the way is
surrounded and carried off, never to be seen again; a lion, who
thought to gain an easy prey, finds himself forced to relinquish his
victim, and to travel with the stream. Unceasingly those behind press
forward, and those in front yield slowly to the pressure; those cooped
up in the middle strive continually to reach the wings, and their
efforts are strenuously resisted. Above the clouds of dust raised by
the rushing army the vultures circle; flanks and rear are attended by
a funeral procession of various beasts of prey; in the passes lurk
sportsmen, who send shot after shot into the throng. So the tortured
animals travel for many miles, till at length spring sets in and their
armies are broken up.
Shall I go on to consider other compulsory migrations, such as those
of the arctic foxes and polar bears when an ice-floe on which they
were hunting is loosened and floated off by the waves till, under
favourable circumstances, it touches some island? I think not, for
journeys such as these are not migrations, they are simply passive
driftings.

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Pharmacology of the WNT Signaling

System 1st Edition Gunnar Schulte

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Pharmacology of the WNT Signaling

ISSN 0171-2004 e-ISSN 1865-0325

© The Editor(s) (if applicable) and The Author(s), under exclusive

The use of general descriptive names, registered names, trademarks,

This Springer imprint is published by the registered company Springer

Controlling Wnt Signaling Specificity

Keywords Engineered Wnt agonist – Extracellular environment –

2 Wnt Processing and Secretion

4 Wnt Signaling Specificity

6 Targeting Wnt in Disease

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