1

CURS OPTIONAL
GENETICA MEDICALA SI
BIOLOGIE MOLECULARA
1866
1944

1953
James Rothman- directorul departamentului de Biologie Celulara de la Yale
(SUA),
Randy Schekman -cercetator la Universitatea California, Berkeley (SUA)
Thomas Sudhof profesor la Universitatea Stanford (SUA),
2013
Human Genome Project
Human Genome Project
Goals:
identify all the approximate 30,000 genes in human DNA,
determine the sequences of the 3 billion chemical base pairs that make up
human DNA,
store this information in databases,
improve tools for data analysis,
transfer related technologies to the private sector, and
address the ethical, legal, and social issues (ELSI) that may arise from the project.

Milestones:
1990: Project initiated as joint effort of U.S. Department of Energy and the National
Institutes of Health
June 2000: Completion of a working draft of the entire human genome
February 2001: Analyses of the working draft are published
April 2003: HGP sequencing is completed and Project is declared finished two years
ahead of schedule
U.S. Department of Energy Genome Programs, Genomics and Its Impact on Science and Society, 2003
What does the draft human
genome sequence tell us?

By the Numbers
The human genome contains 3 billion chemical nucleotide bases (A, C, T, and G).
The average gene consists of 3000 bases, but sizes vary greatly, with the largest
known human gene being dystrophin at 2.4 million bases.
The total number of genes is estimated at around 30,000--much lower than
previous estimates of 80,000 to 140,000.
Almost all (99.9%) nucleotide bases are exactly the same in all people.
The functions are unknown for over 50% of discovered genes.
U.S. Department of Energy Genome Programs, Genomics and Its Impact on Science and Society, 2003
What does the draft human
genome sequence tell us?
The Wheat from the Chaff

Less than 2% of the genome codes for proteins.

Repeated sequences that do not code for proteins ("junk DNA") make up at least
50% of the human genome.

Repetitive sequences are thought to have no direct functions, but they shed light
on chromosome structure and dynamics. Over time, these repeats reshape the
genome by rearranging it, creating entirely new genes, and modifying and
reshuffling existing genes.

The human genome has a much greater portion (50%) of repeat sequences than
the mustard weed (11%), the worm (7%), and the fly (3%).
U.S. Department of Energy Genome Programs, Genomics and Its Impact on Science and Society, 2003
How does the human genome
stack up?
Organism Genome Size
(Bases)

Estimated
Genes
Human (Homo sapiens) 3 billion 30,000
Laboratory mouse (M. musculus) 2.6 billion 30,000
Mustard weed (A. thaliana) 100 million 25,000
Roundworm (C. elegans) 97 million 19,000
Fruit fly (D. melanogaster) 137 million 13,000
Yeast (S. cerevisiae) 12.1 million 6,000
Bacterium (E. coli) 4.6 million 3,200
Human immunodeficiency virus (HIV) 9700 9
Gene number, exact locations, and functions
Gene regulation
DNA sequence organization
Chromosomal structure and organization
Noncoding DNA types, amount, distribution, information content, and
functions
Coordination of gene expression, protein synthesis, and post-translational
events
Interaction of proteins in complex molecular machines
Proteomes (total protein content and function) in organisms
Correlation of SNPs (single-base DNA variations among individuals) with
health and disease
Disease-susceptibility prediction based on gene sequence variation
Genes involved in complex traits and multigene diseases
Developmental genetics, genomics
Future Challenges:
What We Still Dont Know
U.S. Department of Energy Genome Programs, Genomics and Its Impact on Science and Society, 2003
Anticipated Benefits of
Genome Research
Molecular Medicine
improve diagnosis of disease
detect genetic predispositions to disease
create drugs based on molecular information
use gene therapy and control systems as drugs
design custom drugs (pharmacogenomics) based on individual genetic profiles

Microbial Genomics
rapidly detect and treat pathogens (disease-causing microbes) in clinical practice
develop new energy sources (biofuels)
monitor environments to detect pollutants
protect citizenry from biological and chemical warfare
clean up toxic waste safely and efficiently
U.S. Department of Energy Genome Programs, Genomics and Its Impact on Science and Society, 2003
Risk Assessment
evaluate the health risks faced by individuals who may be exposed to radiation
(including low levels in industrial areas) and to cancer-causing chemicals and toxins
Bioarchaeology, Anthropology, Evolution, and Human Migration
study evolution through germline mutations in lineages
study migration of different population groups based on maternal inheritance
study mutations on the Y chromosome to trace lineage and migration of males
compare breakpoints in the evolution of mutations with ages of populations and
historical events
U.S. Department of Energy Genome Programs, Genomics and Its Impact on Science and Society, 2003
Anticipated Benefits of
Genome Research-cont.
DNA Identification (Forensics)
identify potential suspects whose DNA may match evidence left at crime
scenes
exonerate persons wrongly accused of crimes
identify crime and catastrophe victims
establish paternity and other family relationships
identify endangered and protected species as an aid to wildlife officials (could be
used for prosecuting poachers)
detect bacteria and other organisms that may pollute air, water, soil, and food
match organ donors with recipients in transplant programs
authenticate consumables such as caviar and wine

U.S. Department of Energy Genome Programs, Genomics and Its Impact on Science and Society, 2003
Anticipated Benefits of
Genome Research-cont.
Anticipated Benefits:
improved diagnosis of disease
earlier detection of genetic predispositions to disease
rational drug design
gene therapy and control systems for drugs
personalized, custom drugs
Medicine and the New
Genetics
U.S. Department of Energy Genome Programs, Genomics and Its Impact on Science and Society, 2003
Gene Testing Pharmacogenomics Gene
Therapy
ELSI: Ethical, Legal,
and Social Issues
Privacy and confidentiality of genetic information.
Fairness in the use of genetic information by insurers, employers, courts, schools,
adoption agencies, and the military, among others.
Psychological impact, stigmatization, and discrimination due to an individuals
genetic differences.
Reproductive issues including adequate and informed consent and use of genetic
information in reproductive decision making.
Clinical issues including the education of doctors and other health-service providers,
people identified with genetic conditions, and the general public about capabilities,
limitations, and social risks; and implementation of standards and quality-control
measures.
U.S. Department of Energy Genome Programs, Genomics and Its Impact on Science and Society, 2003
HapMap
Begun in 2002, the project is a 3-year effort to construct
a map of the patterns of SNPs (single nucleotide
polymorphisms) that occur across populations in Africa,
Asia, and the United States.
Consortium of researchers from six countries
Researchers hope that dramatically decreasing the
number of individual SNPs to be scanned will provide a
shortcut for identifying the DNA regions associated with
common complex diseases
Map may also be useful in understanding how genetic
variation contributes to responses in environmental
factors
GWAS(genome wide association study) A genome-
wide association study (GWAS), also known as whole
genome association study (WGAS), is an
examination of all or most of the genes (the genome)
of different individuals of a particular species to see
how much the genes vary from individual to
individual
Representative Genomewide Association Studies of Common Cardiovascular Diseases.
O'Donnell CJ, Nabel EG. N Engl J Med 2011;365:2098-2109

GENOMICA / PROTEOMICA / METABOLOMICA
Genomica studiaza genomul cu toate componentele structurale
(acizi nucleici, proteine)

Cercetarea unei singure gene nu intra in cadrul definitiei genomicii,
cu exceptia cazului in care scopul analizei acestei informatii
genetice si functionale este de a elucida efectul si locul sau in cadrul
intregii retele a genomului.

Biologia moleculara se ocupa cu intelegerea interactiunilor dintre
diferite sisteme ale celulei (inclusiv interactiunile dintre ADN, ARN si
biosinteza proteinelor) si deasemenea cu intelegerea modului in
care aceste interactiuni sunt reglate.


MEDICINA GENOMICA
Medicina genomic este o medicin
individualizat, personalizat i predictiv,
preventiv, proactiv (prospectiv) i
participativ (regula celor 5 P).
ONCOGENOMICA
Medicina genomica
ONCOGENETICA
Fiecare tumor conine un numr foarte mare de
modificri genetice i epigenetice diverse.
numai un numr mic de mutaii (mai puin de zece),
numite mutaii conductoare (driver mutations) sunt
necesare pentru iniierea clonei canceroase (mutaii
oncogenice) i pentru meninerea/expansiunea
tumoral (mutaii de meninere)
unele mutaii conductoare au fost definite ca modificri
acionabile (actionable aberrations), n sensul c
implic aciuni practice deoarece au impact asupra
managementului cancerului prin utilizarea lor n
diagnostic, prognostic i/sau predicie;
o parte dintre acestea sunt i tratabile(druggable
aberrations) ntruct oncoproteinele codificate pot fi
inta unor noi terapii, care schimb evoluia bolii
ONCOGENETICA
Inductia apoptozei prin letalitatea sintetica
Acest concept se bazeaz pe
interaciunea genetic dintre dou sau mai
multe gene sau ci moleculare;
inhibarea lor combinat/concomitent
produce eliminarea selectiv a clonelor de
celule premaligne, fr efecte citostatice
asupra celulelor normale i deci fr
efecte toxice.
ONCOGENETICA
Astfel, celulele canceroase dezvolt rezisten la
apoptoz, prin activarea unor ci de
supravieuire celular, secundar unor mutaii
(de exemplu, a genei APC n cancerul colorectal
sau a oncogenei KRAS n cancerul pulmonar
fr celule mici);

celulele canceroase cu aceste mutaii pot fi
eliminate prin folosirea unei combinaii sintetic-
letale
ONCOGENETICA
Un alt exemplu de letalitate sintetic este reprezentat de
celulele canceroase cu mutaii ale genelor de reparare
ale leziunilor ADN (de tipul BRCA1 i BRCA2 n cancerul
ereditar de sn i ovar sau PTEN n cancerul de
prostat).

Administrarea unor enzime implicate n repararea ADN,
duce la cumularea efectelor (amplificarea rupturilor ADN)
n celulele canceroase i moartea acestora) efect
coroborat cu radioterapia.

Conceptul letalitii sintetice poate fi utilizat pentru o
terapie personalizat n cancer, determinat de profilul
individual oncogen
ONCOGENETICA
leziunile genetice gsite n trunchiul principal
(clona iniial) sunt exprimate i n ramurile
sale (subclone).

n aceste condiii, se impune schimbarea
strategiilor terapeutice i direcionarea lor spre
gen i nu spre proteina codificat de gen,
urmrind inhibiia ei funcional sau restaurarea
structurii ei normale

PROTEOMICA
Proteomica se ocupa cu studiul la scara larga a structurii si functiei proteinelor.

Analiza de secven poate furniza mai multe date prin compararea secvenei
unei proteine cu cele stocate n bazele de date.
se pot gsi informaii despre: structura, interaciile, activitatea biochimic,
evoluia i chiar rolul potenial ntr-o boal.


Structura tridimensional depinde de plierea catenei polipeptidice n spaiu care
reflect:
Lungimea secvenei (tip de aminoacizi i secven)
Structura primar
Modificri post translaionale suferite de aminoacizi


Proteinele cu secvene similare au proprieti similare. Paradigma
secven similar/ funcie similar st la baza bioinformaticii i ne permite
s se fac predicii structurale i funcionale pe baza secvenei proteice.
T E H N O L O G I A P C R
Reacia PCR (Polymerase Chain Reaction = Reacie de Polimerizare n
Lan) este o metod de amplificare enzimatic in vitro a unei anumite
secvene de ADN.
tehnica PCR permite ca o singura copie a unei secvente de ADN sa fie
copiata de milioane de ori sau chiar sa fie modificata pe parcursul
amplificarii (atunci cand este dorit acest lucru: ex. Introducerea unei
mutatii).
Din punct de vedere chimic, reacia PCR este constituit din cicluri succesive
de replicare ADN in vitro, folosind 2 primeri oligonucleotidici ce hibridizeaz
cu cele 2 catene ale secvenei originale (folosit ca matri n replicare).

Diferena esenial ntre o asemenea reacie de replicare i un proces de
replicare ADN in vivo, l reprezint faptul c n reacia PCR etapa de
desfacere a dublului helix matri i, respectiv, cea de ataare a primerilor, nu
sunt realizate enzimatic, ci prin parcurgerea unor trepte de temperatur, iar
singura enzim folosit n reacie este o ADN polimeraz ADN-dependent
(cu funcie de replicaz).


TIPURI DE PCR
Real Time PCR (PCR cantitativ),
Real Time PCR este o metoda diferita
fata de RT-PCR (reverse transcriptase
PCR).
Real Time PCR este o metoda care
include atat amplificarea fragmentului
ADN in timp real cat si analiza acestuia.
Se folosesc cantitati foarte mici de
proba. Se pot amplifica atat probe ADN
cat si probe ADNc cu aceeasi viteza si
eficienta
Un numar mult mai mare de probe de
concentratii diferite pot fi analizate in
acelasi timp.
Amplificarea fragmentului ADN se
poate vizualiza cu ajutorul moleculei
reporter pe tot parcursul reactiei, nu la
sfarsit.
Aplicatii Real-Time PCR
Real-Time PCR poate fi utilizat pentru:
Genotipare
Trisomii si detectarea numarului de gene unice din genom
Genotiparea microdeletiilor
Haplotipare
Analiza cantitativa a microsatelitilor
Diagnosticul prenatal al celulelor fetale din sangele matern
Diagnosticul cancerelor
Cuantificarea expresiei genelor
Masurarea expunerii la radiatii
Studiul ADN mitocondrial
Detectia metilarii
Detectia inactivarii cromosomului X

Clonarea moleculara si Expresia genelor

Clonarea molecular se refer la procedura de izolare a unei anumite
secvene ADN i obinerea mai multor copii ale acesteia in vitro.
Clonarea este frecvent folosit pentru amplificarea secvenelor ADN ce
conin gene, dar poate fi folosit i pentru amplificarea oricrei secvene
ADN cum ar fi promotorii, secvene necodate i secvene aleatoare ale ADN.
Enzimele de restrictie sunt o clasa speciala de enzime utilizate pentru a taia
ADN-ul la un anumit site de restrictie pe care il recunosc, avand ca rezultat
fragmente de ADN de lungimi predictibile.
Utilizarea enzimelor de restrictie permite ca fragmente de ADN din surse
diferite sa fie reconectate si astfel, prin ligarea lor, este obtinut ADN-ul
recombinat.

Adesea asociat cu organismele modificate genetic, ADN-ul recombinat este
utilizat in constructia vectorilor care au la baza plasmidele (fragmente scurte
de ADN circular).
Clonarea moleculara si expresia genelor
Vectorii de expresie sunt un tip specializat de vectori de clonare in
care semnalele pentru translatie si transcriptie necesare pentru
controlul genei de interes sunt incluse in vectorul de clonare. Aceste
semnale pot fi create artificial pentru a controla mai usor expresia
genei de interes.

Expresia genelor este o tehnica de clonare ADN care utilizeaza
vectori de clonare pentru a genera o biblioteca de clone, in care
fiecare clona exprima o proteina. Biblioteca de expresie este
analizata pentru identificarea clonei care intereseaza si aceasta este
recuperata pentru investigatii ulterioare. Un exemplu ar fi izolarea
unei gene care confera rezistenta la antibiotice.

Scop De obicei scopul final al expresiei genelor este a produce o
anumita proteina in cantitate mare.

EXPRESIA GENELOR -APLICATII
Obinerea insulinei umane.
Noile tehnologii industriale de obinere a insulinei umane au fost
posibile odat cu extragerea genei insulinei (W.Gillbert i
colaboratorii si, 1980) i crearea moleculelor recombinate de ADN
n baza plasmidelor

Moleculele recombinate de ADN sunt transferate n Escherichia
coli, unde are loc realizarea informaiei genetice codificate n
molecula de ADN. Paralel cu proteinele specifice bacteriei, se
sintetizeaz i insulina. Pentru a proteja insulina uman n molecula
recombinat de ADN se ncadreaz, pe lng gena insulinei, i o
gen reglatoare care codific proteine specifice colibacililor
(galactozidaza). Ca rezultat al manifestrii informaiei genetice a
moleculei recombinate de ADN, se obine o caten polipeptidic
hibrid, din care mai apoi se separ insulina.

se obin aproximativ 200 grame de insulin de pe 1 m
3
de mediu de
cultur (1600 kg de pancreas de bou sau porc.)
Obinerea somatotropinei.

Sinteza acestui hormon pe cale artificial s-a nceput cu producerea
de ADNc cu ajutorul revers-transcriptazei, avnd ca matrice
ARNm din hipofize (transcripie invers).

Acesta a fost clonat, apoi tiat cu enzime de restricie pentru
obinerea secvenei nucleotidice corespunztoare somatotropinei,
cu excepia fragmentului ce determin primii 23 de aminoacizi.

Fragmentul n cauz era clonat separat, ca rezultat al unei sinteze
chimice, apoi cele dou segmente unite, la ele se adug segmente
reglatoare i pe baza plasmidelor se obine plasmidiul recombinat
cu gena HGH (a somatotropinei).

Colibacilii, primind acest plasmid recombinat, sintetizau
somatotropina (la 1 litru de mediu de cultur se obine 2,0-2,5 mg
somatotropin).

EXPRESIA GENELOR- APLICATII


TELOCIT

Telocitele sunt celule cu corp mic, dar cu prelungiri
extrem de lungi (asemanatoare prelungirilor neuronilor),
prezente in aproape toate organele.

Rolul telocitelor este acela de tele-coordonare, la
distanta, prin intermediul prelungirilor lor extrem de lungi
si subtiri (telopode), a altor tipuri celulare, de stabilire de
contacte de tip "sinapse stromale" cu acestea si de
comunicare prin intermediul unor structuri
microveziculare - exozomi

Se folosesc in terapia regeneratoare post-infarct de
miocard
NIGMS Image Gallery


Prima data termenul EPIGENETICA a fost utilizat pentru a
explica trasaturi fenotipice care nu puteau fi justificate prin
mecanisme genetice (Conrad Waddington 19051975)

Epigenetica promite sa descifreze procesele biologice implicate in
dezvoltare si diferentiere, interfata dintre organisme si
perturbarile induse de mediu (inclusiv carcinogeneza)


Epigenetica = ansamblul de mecanisme care moduleaza
structura cromatinei si regleaza stabilitatea expresiei genice
pentru definirea identitatii celulare







1. Desi majoritatea celulelor umane au aceeasi
informatie genetica cuprinsa in ADN, organismul
produce in cursul diferentierii diferite tipuri de celule;

2. Fiecare celula are profilul propriu de expresie genica
si functii specifice.

3. Exista unui numar mare de studii care arata ca
genetica (singura!) nu poate explica toate aspectele
legate de dezvoltare si diferentiere

4. Importanta factorilor non-genetici sustinuta inclusiv
de observatii la unele animale clonate, probabil si ca
rezultat al expresiei incorecte in imprinting.


Informatia epigenetica determina modularea
expresiei genice conform unui pattern determinat
de etapa de dezvoltare, de tipul de tesut si de
conditile ambientale determinind un anumit fenotip
celular.

Replicarea cromatinei in cursul fazei S a ciclului
celular ofera oportunitatea factorilor responsabili sa
adauge si sa propage in noua catena ADN toti
markerii epigenetici identificati in celula parentala.

Epigenetica a adus rezultate remarcabile in
optimizarea strategiilor de diagnostic (diagnosticul
timpuriu si preventia) si in elaborarea strategiilor
terapeutice epigenetice.
EPIGENETICA
Primul mecanism epigenetic descris este
inactivarea cromozomului X(teoria Mary
Lyon/ metilare)


.
1. Metilarea ADN
-Adaugarea covalenta a unei grupari metil (-CH
3
) in pozitia 5 a
citozinei cu obtinerea 5-metil citozinei (5mC);





5mC reprezinta 1% din totalul bazelor azotate

-Donor de grupari metil: S-adenozil metionina (SAM)

-Situsuri preferentiale de metilare: secvente CpG

-Originea grupelor metil: dieta (metabolismul folatului sau
metioninei, seleniu)







Nucleotidele CpG
Distribuite in intreg genomul
Concentrate in regiuni numite insule CpG
(60% din ADN genomic)

Localizate preferential la nivelul:
- promotorului genelor
- regiunilor ADN reglatoare
-ADN repetitiv (palindroame, ADN satelit)

clusterele CpG nemetilate sunt situate in genele
implicate in desfasurarea optima a activitatii
celulare (ex. gene housekeeping).

Pattern-ul de metilare
Conservat in celulele somatice
Realizat de ADN metiltransferaze (DNMT- DNA
methyltransferases)
DNMT1- actioneaza preferential asupra ADN hemi-
metilat.
-metileaza catenele nou sintetizate
imediat dupa replicare, folosind matrita
parentala.

ADN metiltransferaze de novo:
- DNMT3A
- DNMT3B
. Tumorigenic Mechanisms in Mammalian Cells. Both genetic and
epigenetic aberrations are involved in neoplastic transformation. These 2
alternate pathways of tumorigenesis are linked by an intricate cross-talk
and can, either individually or in synergy, lead to the development of the
malignant phenotype
Metilarea aberanta ADN si cancer
Defectele de metilare se pot transmite pe
linie germinala- cancere familiale (colon,
endometru)
Metilarea genei p16 in leziuni neoplazice
la fumatori
(dg. In sputa cu sensibilitate 96%) pentru
cancerul pulmonar
lipsa imprinting Insulin like growth
factor in neoplasmul colorectal (dg. materii
fecale)
. Folate metabolism: folate, along with choline, methionine, cobalamin, pyridoxine, and
riboflavin, is involved in several essential metabolic processes within the cell, in particular
DNA synthesis, repair, and methylation. Folate is also essential as a methyl donor
INFLUENTA FOLATULUI IN METILAREA
ADN
Figure 4. Possible mechanisms by which folate deficiency or excess
may influence telomere structure and function: indentifies
plausible but untested mechanisms.
2. Structura cromatinei
Cromatina- organizata in 2 nivele:
- heterocromatina silentiata
(telomere, regiuni pericentrice,
secvente repetitive)

-eucromatina activa (genele active)

Nucleosom = unitatea de baza a
cromatinei (ADN dublu catenar
infasurat in jurul octamerului histonic).

Miezul histonic - 2 copii ale
fiecarei histone (H2A, H2B,H3 si H4)
Lodish, 2000

Caracteristicile modificarilor post-translationale ale
histonelor

-sunt modificari chimice, covalente, dinamice si reversibile;
-nu sunt aleatoare;
-realizate de enzime
- factorii de initiere:
-semnale endogene - etapa de dezvoltare
- tipul de tesut
- semnale exogene - factori de mediu

Rolul: - afecteaza organizarea secundara a cromatinei
- realizeaza interactii intre ADN si histone




Modelare
expresie
genica
fenotip
Tipuri de modificari post-translationale ale
histonelor
Modificarile se realizeaza la aminoacizii din capatul N-
terminal al histonelor



Dupa Tomasi TB et al, 2006
Modularea transcrierii
Acetilarea si metilarea sunt considerate modulatorii
cheie ai activarii si represiei transcriptionale


Rodenhiser D, Mann M, 2006
activ activ
Adaptat dupa Zelent A
et al, 2005
Codul epigenetic
Mecanism metilare oncogen
Fig. 2. (a), aberrant methylation keeps occurring. After several clonal selections during multistep
carcinogenesis, all cancer cells come to have methylation of multiple CGI. In contrast, if a cancer cell is
derived from a precursor cell with methylation of multiple CGI (b), cancer cells
derived from it will also display methylation of multiple CGI.
CELULE STEM REPROGRAMATE
John B. Gurdon a descoperit n 1962 faptul c specializarea celulelor este
reversibil.

n cadrul unui experiment devenit clasic, el a nlocuit nucleul celular imatur
dintr-o celul ou (embrionar) de broasc cu nucleul prelevat de la o celul
intestinal matur.
Celula embrionar modificat s-a dezvoltat ntr-un mormoloc de broasc
normal.
El a demonstrat c ADN-ul celulei mature, dei specializate, dispune
de toate informaiile necesare pentru dezvoltarea tuturor celorlalte
celule specializate care alctuiesc organismul broatei.



Shinya Yamanaka reuseste s reprogrameze celule mature de oareci
pentru a deveni celule stem (imature, nespecializate).

n mod surprinztor, prin introducerea a doar ctorva gene, el a reuit s
reprogrameze celulele mature n celule stem pluripotente - celule
imature, capabile s se specializeze n orice tip de celul a
organismului.


CELULE STEM PLURIPOTENTE INDUSE SI
SUPRAEXPRESIA FACTORILOR DE TRANSCRIPTIE
3. ARN necodificator (ncRNA)

Cel mai studiat ARNnc este microARN

microARN (miRNA)

Molecule mici de ARN endogen (22 nucleotide),
monocatenar, necodificator
Se leaga la capatul 3 (3UTR) al unui/unor ARN
mesager specific(i)
Moduleaza expresia genica prin
- represarea expresia proteinei tinta
- degradarea ARNm specific
Poate controla direct structura cromatinei tintind factori
implicati in remodelarea acesteia (conexiune cu PcG)
In imunologie, miRNA sunt considerati reglatori cheie
in angajarea pe o linie de diferentiere, maturare,
mentinerea homeostaziei

The Increasing Complexity of the Central Dogma of Molecular Biology.
Feero WG et al. N Engl J Med 2010;362:2001-2011.
Biogeneza/ actiune
Istoria miRNA
miRNA in hepatopatii
EXOZOM
Exozomii au markeri de suprafata CD63, CD9, CD81.

Deriva din linie hematopoetica :reticulocite, trombocite si
limfocite .
Pot fi secretati si de celule tumorale epiteliale ( neo
pulmonar, renal, intestinal).

Studii recente au demonstrat ca pot transporta mRNA si
microRNA.

Datorita diversitatii de transport au fost asociati cu
mecanismele de polarizare a celulelor epiteliale,
neuronale si crestere tumorala.

In studiile clinice se evalueaza nivelul exozomilor in
sange, plasma, urina, lichid amniotic si lichide
neoplazice.