# Targeting DNA Demethylation Regulators in Osteoarthritis

> **NIH NIH R01** · STANFORD UNIVERSITY · 2020 · $347,600

## Abstract

Abstract
Osteoarthritis (OA) is a complex age-associated disorder with an unidentified etiology. Our long-term goal is to
understand the epigenetic mechanisms underlying OA pathology especially the role of DNA methylation and
demethylation. Conversion of methyl cytosine (5mC) to its hydroxylated form (5hmC), catalyzed by the ten-
eleven translocation (TET) enzymes acts as an intermediate in active DNA demethylation. We have recently
reported that OA patients have a dysregulation of the 5hmC homeostasis in cartilage, leading to increased
5hmC levels that are associated with activated OA genes. Our new unpublished findings show that TET1
knockout mice are resistant to OA development and that the key OA genes MMPs 3 and 13 are potential
targets of TET1 and TET2. Based on these findings, we propose to test the central hypothesis that TET
proteins-mediated DNA modifications activate OA-associated genes and that loss of TET function can be
therapeutic in OA. Firstly, we will determine how expression of OA-associated genes is activated by 5hmC
enrichment and DNA demethylation in human OA chondrocytes. We will study the global distribution of 5hmC
and 5mC as well as gene expression in a cohort of non-OA and OA chondrocytes, using state-of-the-art
chemical labeling and enrichment techniques followed by high-throughput sequencing (hme-Seal, MBD-seq
and RNA-seq respectively). Secondly, we will directly effect a specific loss of TET1/2 in human chondrocytes
and in post-natal cartilage in `conditional' KO mice. Treatment with inflammatory cytokines will be utilized in
vitro in the presence or absence of TET function to reveal direct TET targets and their precise mode of
regulation. For the in vivo studies, we will utilize OA induction in TET1 and TET2 conditional knockout mice to
determine the effect of TET loss in the early and late stages of OA pathology. The major outcomes of these
studies will be to (a) identify OA-associated target genes regulated by TET1 and 2, (b) identify 5mC and 5hmC
dependent gene expression changes in early and late stages of OA and (c) and most importantly identify how
TET inhibition can be utilized to modulate OA pathogenesis. Collectively, these studies have the potential to
shed light on a new facet of OA pathogenesis and to identify new therapeutic strategies for modifying OA.

## Key facts

- **NIH application ID:** 9979766
- **Project number:** 5R01AR070865-05
- **Recipient organization:** STANFORD UNIVERSITY
- **Principal Investigator:** Nidhi Bhutani
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $347,600
- **Award type:** 5
- **Project period:** 2016-09-07 → 2022-12-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9979766

## Citation

> US National Institutes of Health, RePORTER application 9979766, Targeting DNA Demethylation Regulators in Osteoarthritis (5R01AR070865-05). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/9979766. Licensed CC0.

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