LECTURE

INTRACELLULAR ACCUMULATION AND

CELLULAR AGING

Masum Shahriar
Professor, Department of Pharmaceutical Sciences

Reading Assignment
Robbins: Pathologic Basis of Disease, Saunders, 9th edition,
Chapter 1 (ISBN 0-7216-0187-1)

Learning Objectives:
At the end of this lecture, candidates should be able to:
* Describe the cellular accumulations that commonly occur in disease
and with stress, and describe their etiology and pathophysiology
* Define aging and differentiate from normal development and disease
* Describe the morphologic changes that are associated with cellular
aging
* Describe factors (such as genetic, dietary, and disease) influence the
aging process
* Define hypotheses relating cellular metabolic damage to cell aging and
lifespan.
* Describe the proposed role of aerobic metabolism to cellular aging

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 AUTOPHAGY

Autophagy (“self-eating”) refers to lysosomal

digestion of the cell’s own components. It is a
survival mechanism in times of nutrient
deprivation, such that the starved cell subsists by
eating its own contents and recycling these
contents to provide nutrients and energy.

Cellular stresses, such as nutrient deprivation, activate autophagy

genes (Atg genes), which initiate the formation of autophagic
vacuole in which cellular organelles are sequestered. Autophagic
vacuole is postulated to be formed from ribosome-free regions of
the ER. The vacuole fuses with lysosomes to form an
autophagolysosome, in which lysosomal enzymes digest the cellular
components and the products are used to provide nutrients for the
cell. With time, the starved cell eventually can no longer cope by
devouring itself; at this stage, autophagy may also signal cell death
by apoptosis.

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 INTRACELLULAR ACCUMULATIONS

Under some circumstances cells may accumulate

abnormal amounts of various substances, which
may be harmless or associated with varying
degrees of injury. The substance may be located in
the cytoplasm, within organelles (typically
lysosomes), or in the nucleus, and it may be
synthesized by the affected cells or may be
produced elsewhere.

Intracellular Accumulations
There are four main pathways of abnormal
intracellular accumulations:
• Inadequate removal of a normal substance
secondary to defects in mechanisms of packaging
and transport, as in fatty change in the liver
• Accumulation of an abnormal endogenous
substance as a result of genetic or acquired defects
in its folding, packaging, transport, or secretion, as
with certain mutated forms of α1-antitrypsin.
• Failure to degrade a metabolite due to inherited
enzyme deficiencies. The resulting disorders are
called storage diseases.
• Deposition and accumulation of an abnormal
exogenous substance when the cell has neither the
enzymatic machinery to degrade the substance nor
the ability to transport it to other sites.
Accumulation of carbon or silica particles is an
example of this type of alteration.

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• Depositions of lipids
Fatty change: accumulation of free triglycerides in cells,
resulting from excessive intake or defective transport (often
because of defects in synthesis of transport proteins);
manifestation of reversible cell injury
Cholesterol deposition: result of defective catabolism and
excessive intake; in macrophages and smooth muscle cells of
vessel walls in atherosclerosis
• Deposition of proteins: reabsorbed proteins in kidney
tubules; immunoglobulins in plasma cells
• Deposition of glycogen: in macrophages of patients with
defects in lysosomal enzymes that break down
glycogen(glycogen storage diseases)
• Deposition of pigments: typically indigestible pigments, such
as carbon, lipofuscin (breakdown product of lipid peroxidation),
or iron (usually due to overload, as in hemosiderosis)

PATHOLOGIC CALCIFICATION

Pathologic calcification is a common process in a wide

variety of disease states; it implies the abnormal deposition
of calcium salts, together with smaller amounts of iron,
magnesium, and other minerals. Calcification may be
classified into

 dystrophic calcification: When the deposition occurs in dead or

dying tissues, it is called dystrophic calcification; it occurs in the
absence of derangements in calcium metabolism (i.e., with normal
serum levels of calcium).

 metastatic calcification: the deposition of calcium salts in normal

tissues is known as metastatic calcification and is almost always
secondary to some derangement in calcium metabolism
(hypercalcemia).

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 Regardless of the site, calcium salts are seen on gross
examination as fine white granules or clumps, often felt as gritty
deposits. Dystrophic calcification is common in areas of caseous
necrosis in tuberculosis. Sometimes a tuberculous lymph node is
essentially converted to radiopaque stone. On histologic
examination, calcification appears as intracellular and/or
extracellular basophilic deposits. Over time, heterotopic bone may
be formed in the focus of calcification.

Metastatic calcification can occur widely throughout the body but

principally affects the interstitial tissues of the vasculature, kidneys,
lungs, and gastric mucosa. The calcium deposits morphologically
resemble those described in dystrophic calcification. Although they
generally do not cause clinical dysfunction, extensive calcifications
in the lungs may be evident on radiographs and may produce
respiratory deficits, and massive deposits in the kidney
(nephrocalcinosis) can lead to renal damage.

The major causes of hypercalcemia are

(1) increased secretion of parathyroid hormone, due to either
primary parathyroid tumors or production of parathyroid
hormone–related protein by other malignant tumors;
(2) destruction of bone due to the effects of accelerated
turnover (e.g., Paget disease), immobilization, or tumors
(increased bone catabolism associated with multiple
myeloma, leukemia, or diffuse skeletal metastases);
(3) vitamin D–related disorders including vitamin D
intoxication and sarcoidosis (in which macrophages
activate a vitamin D precursor); and
(4) renal failure, in which phosphate retention leads to
secondary hyperparathyroidism.

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 Cellular Aging
Cellular aging is the result of a progressive decline in the
life span and functional capacity of cells.

Aging is a continuum of processes that begin with conception,

involves differentiation and maturation, and at some point leads
to progressive loss of functional capacity characteristic of
senescence and ends in death.

Most people would classify "aging changes " as relating to

those that occur after maturation of the organism.

Characteristic changes of aging in organ systems

Skeletal system - Loss of bone mass, often reduced stature

Skin - Wrinkles and sagging

Heart, Kidney and Lungs - Often reduced reserve capacity

Brain - Loss of some neurons

Body fat – Redistribution

Hormones - Changes in secretion pattern of hormones

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 Aging in individuals is affected by:

* Genetic factors - some genetic variants undergo

premature aging

* Diet - Dietary restriction can prolong life span (rodents,

primates)

* Disease - Some diseases may accelerate the "aging"

process (e.g., diabetes)

Mechanisms of cellular aging

Several mechanisms are thought to be responsible for cellular
aging

Mechanisms that cause and counteract cellular aging.

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 DNA damage

A variety of metabolic insults that accumulate over time may

result in damage to nuclear and mitochondrial DNA.
Although most DNA damage is repaired by DNA repair
enzymes, some persists and accumulates as cells age.
Some aging syndromes are associated with defects in DNA
repair mechanisms, and the life span of experimental
animals in some models can be increased if responses to
DNA damage are enhanced or proteins that stabilize DNA
are introduced.

Decreased cellular replication.

All normal cells have a limited capacity for replication, and

after a fixed number of divisions cells become arrested in a
terminally nondividing state, known as replicative
senescence. Aging is associated with progressive
replicative senescence of cells. Cells from children have
the capacity to undergo more rounds of replication than do
cells from older people. In human cells, the mechanism of
replicative senescence involves progressive shortening of
telomeres, which ultimately results in cell cycle arrest.

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 The role of telomeres and telomerase
Telomeres are short, repeated sequences
of DNA (TTAGGG)
1) Ensure complete replication of
chromosome ends
2) Protect chromosomal ends from
fusion and degradation
When somatic cells replicate, a small
section of the telomere is not duplicated
and telomeres become progressively © Elsevier 2004

shortened. As the telomeres become

shorter, the ends of chromosomes cannot Telomerase directs RNA
be protected and are seen as broken template-dependent DNA
DNA, which signals cell cycle arrest. synthesis, in which
Telomere length is maintained by nucleotides are added to one
nucleotide addition mediated by an strand at the end of a
enzyme called telomerase. chromosome

Telomere-telomerase hypothesis

• In normal somatic cells, there is

no telomerase activity. Therefore,
as most somatic cells age their
telomeres become shorter and
they exit the cell cycle, resulting
in an inability to generate new
cells to replace damaged ones.
• Germ cells and stem cells both
contain active telomerase
• Only the germ cells have
sufficient levels of the enzyme to
stabilize telomere length
completely
• Telomerase activation in cancer
cells inactivates the teleomeric
clock

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 Defective protein homeostasis.

Over time, cells are unable to maintain normal protein

homeostasis, because of increased turnover and decreased
synthesis caused by reduced translation of proteins and
defective activity of chaperones (which promote normal
protein folding), proteasomes (which destroy misfolded
proteins) and repair enzymes. Abnormal protein
homeostasis can have many effects on cell survival,
replication, and functions. In addition, it may lead to
accumulation of misfolded proteins, which can trigger
pathways of apoptosis.

Mechanisms of protein folding and the role of

chaperones

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* Morphologic changes that are characteristic
of cellular aging

• Irregular or abnormally lobed nuclei

• Vacuolated mitochondria

• Decreased endoplasmic reticulum

• Distorted Golgi apparatus

* Altered metabolic functions

that are characteristic of cellular aging

• Increased DNA damage

• Decreased chromosomal damage repair
• Reduced oxidative phosphorylation by mitochondria
• Decreased capacity for nutrients uptake
• Steady accumulation of the pigment lipofuscin (product of lipid
peroxidation and evidence of oxidative damage)
• Accumulation of advanced glycation end products result from
nonenzymatic glycation (capable of crosslinking proteins)
• Abnormally folded proteins

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 Metabolic Events and Genetic Damage

Life span may be influenced by the balance between cellular

damage from metabolic events occurring within the cell and
counteracting repair processes.

One group of metabolic products that are important in this regard is the
"reactive oxygen metabolites” (hydrogen peroxide, superoxide anion and
nitric oxide). These products and their secondary reactive oxygen
metabolities (peroxynitrites) interact with proteins, lipids and nucleic acids
cause reversible and irreversible oxidative modifications.

Total life span is fixed by total metabolic consumption over a lifetime. Smaller
animals generally have greater metabolic rates and shortened life spans compared to
larger animals

* Metabolic Events and Genetic Damage

1) Restriction of caloric intake lowers steady state oxidative damage, slows age
related changes, and extends life span
2) Variation in longevity among species is inversely correlated with rates of
mitochondrial generation of superoxide anion radical
3) In experimental studies overexpression of antioxidative enzymes (superoxide
dismutase) extends life span in certain species
– One gene that helps control life span in worms (c-elegans) encodes
for an insulin receptor (daf2-gene)

Aging may be the cumulative damage that is generated by toxic products of

metabolism.

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