# Project 3: RAGE/DIAPH1 interactions and cellular stress

> **NIH NIH P01** · NEW YORK UNIVERSITY SCHOOL OF MEDICINE · 2021 · $337,794

## Abstract

Project Summary: Project 3
Extensive research has provided strong support for the role of the multi-ligand receptor for advanced glycation
endproducts (RAGE) in the pathogenesis of diabetes complications, such as accelerated cardiovascular (CVD)
and peripheral arterial disease (PAD) and their common complication, ischemia-reperfusion (IR) injury. Due to
its central role in various disease states, antagonism of RAGE signaling pathways is considered to be a
promising therapeutic approach. Our Program Project team's research identified DIAPH1 as an intracellular
effector of RAGE signaling and demonstrated a direct interaction between the intracellular domain (“tail”) of
RAGE, or ctRAGE, and the FH1 (formin homology 1) domain of DIAPH1, in cell types that are relevant to RAGE-
dependent pathologies using both in vitro and in vivo approaches. In macrophages, cardiomyocytes and
endothelial cells, endoplasmic reticulum (ER) and mitochondrial stress, which are part of RAGE-dependent
pathologies, have been linked to the interaction of DIAPH1 with Mitofusin2 (MFN2). Preliminary results from our
labs strongly support the notion that small molecules can inhibit the RAGE-DIAPH1-MFN2 interaction. The
critical question is how this inhibition affects physiologically important interactions of RAGE. The goal of this
Project is to establish the biophysical and structural biology basis for RAGE-DIAPH1 signal transduction and
identify modes of inhibition. We hypothesize that the in vitro mode of RAGE-DIAPH1 inhibition, as revealed by a
combination of biophysical and structural biology tools, such as NMR spectroscopy, will provide a base for
understanding cellular mechanisms of RAGE signaling inhibition and thus can be effectively used to optimize
small molecule inhibitors of RAGE-DIAPH1-MFN2 interactions. NMR spectroscopy will be used as a main
structural tool since ctRAGE and the FH1 domain of DIAPH1, as well as large segments of MFN2, are flexible
and may not be amenable to crystallization or electron cryomicroscopy. We will probe the structural biology of
novel small molecule antagonists of RAGE-DIAPH1 and use in-cell fluorescence techniques to meticulously
characterize the RAGE-DIAPH1-MFN2 interaction. Our Project will work closely with Projects 1 and 2 to explore
the mechanistic basis of this interaction and to identify novel and potent therapeutic strategies and agents for
diabetes, CVD and PAD.

## Key facts

- **NIH application ID:** 10191023
- **Project number:** 5P01HL146367-03
- **Recipient organization:** NEW YORK UNIVERSITY SCHOOL OF MEDICINE
- **Principal Investigator:** ALEXANDER SHEKHTMAN
- **Activity code:** P01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $337,794
- **Award type:** 5
- **Project period:** 2019-08-01 → 2024-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10191023

## Citation

> US National Institutes of Health, RePORTER application 10191023, Project 3: RAGE/DIAPH1 interactions and cellular stress (5P01HL146367-03). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10191023. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*