# 5-AZACYTIDINE INDUCED DIFFERENTIATION

> **NIH NIH R01** · UNIVERSITY OF SOUTHERN CALIFORNIA · 1988 · $153,330

## Abstract

The long term goal of this project is to utilize ananucleoside analogs of
cytidine to understand the role of DNA methylation in cell
differentiation.  We have previously shown that 5-azacytidine (5-aza-CR)
and 5-azadeoxycytidine (5-aza-CdR) induce profound changes in the
differentiated state of mouse C3H 10T1/2 C18 cells and have attributed
these changes to the abilities of the analogs to perturb DNA methylation.
We now want to undertake more detailed studies on the mechanisms of
5-azanucleoside action and to use clonal derivative of 10T1/2 cells treated
with these agents to understand aspects of de novo methylation and the
molecular control of cell differentiation.

The mechanism of cytotoxicity by 5-aza-CdR will be investigated in a series
of clonal derivatives derived after multiple treatments of 10T1/2 cells
with the drug.  These cells are markedly resistant to the cytotoxic
activity of 5-aza-CdR and will be defined with respect to gene
amplification, 5-aza-CdR incorporation and the interaction of incorporated
5-azacytosine with nuclear proteins.  The cells have markedly reduced
levels of 5-methylcytosine (5mC) and undergo consideragle de novo
methylation when dividing in the absence of further drug treatment.  We
will define pathways for the establishment of new methylation patterns
within the cells at the level of specific gene sequences.  we will also
attempt to experimentally raise DNA 5mC levels within cells by treatment
with inhibitors of DNA synthesis so that DNA hypermethylation can be
modulated and genes may be extinguished within cells.  A major part of the
proposal is devoted to themolecular and cellular characterization of
myogenic derivatives derived from 10T1/2 cells by 5-aza-CR treatment.  We
will attempt to isolate genes specific for the commitment phase of myogenic
differentiation within these cells and define their methylation status and
control.  The hypothesis being tested here is that 5-aza-CR induces the
undermethylation of one or a small number of genes whose subsequent
expression defines the myogenic phenotype.

Overall, these studies will increase our knowledge of the mechanisms of
5-aza-CdR action, define mechanisms for de novo methylation within cells
and characterize regulatory genes in differentiating mouse cells.

## Key facts

- **NIH application ID:** 3179259
- **Project number:** 5R01CA039913-05
- **Recipient organization:** UNIVERSITY OF SOUTHERN CALIFORNIA
- **SAM.gov UEI:** G88KLJR3KYT5
- **Principal Investigator:** PETER A JONES
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1988
- **Award amount:** $153,330
- **Award type:** 5
- **Project period:** 1984-09-01 → 1989-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3179259

## Citation

> US National Institutes of Health, RePORTER application 3179259, 5-AZACYTIDINE INDUCED DIFFERENTIATION (5R01CA039913-05). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3179259. Licensed CC0.

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