# Epigenetic modulation of cellular aging to improve CAR-T therapy for solid tumors

> **NIH NIH R43** · DORIAN THERAPEUTICS, INC. · 2021 · $335,648

## Abstract

Abstract
CAR-T cell therapy has had incredible clinical success in the treatment of hematological malignancies. However,
very limited activity against solid tumors has been achieved so far, despite targeting a variety of antigens and
tumor types. Importantly, solid tumors account for more than 90% of all cancers, affecting more than 16 million
Americans in 2020. The goal of Dorian Therapeutic’s SBIR proposal is to investigate the impact of reducing
cellular senescence in CAR-T cells to improve solid tumor targeting.
We have proven that T cells age the equivalent of 30 years during CAR-T manufacturing, which is detrimental
to their ability to expand and kill cancer cells. Moreover, solid tumors are characterized by a strong
immunosuppressive and pro-senescent microenvironment, strongly challenging T cells effector functions. Old T
cells are particularly prone to be suppressed and die due to the hostile microenvironment. Rejuvenating T cells
by reducing cellular aging at the epigenetic level is a novel strategy to improve T cell fitness and clinical
outcomes. The goal of this project is to identify a shRNA-based genetic cassette targeting the epigenetic
regulator USP16 to increase CAR-T cell fitness for the treatment of solid tumors.
Dorian’s founders identified USP16 at Stanford University as an epigenetic regulator able to reduce senescence
by enhancing stem cell self-renewal in multiple tissues. USP16 is a deubiquitinating enzyme responsible for the
removal of ubiquitin moieties from histone H2AK119, increasing chromatin accessibility to pro-senescent
programs. The company has developed in vitro and in vivo proof-of-concept that targeting USP16 by means of
a shRNA co-expressed within the CD19.CAR construct reduces T cell aging and increases stem cell memory
(Tscm) frequency upon manufacturing, without affecting proliferation. Most importantly, the engineered CAR-T
cells resulted in improved tumor killing in a preclinical mouse model of leukemia.
The Aims of this proposal are to (1) Create an in vitro model to study the effect of tumor microenvironment on T
cells aging and functions, (2) Identify genetic cassettes targeting USP16 and increasing T cell fitness in vitro,
and (3) Demonstrate that engineered CAR-T cells expressing a shRNA for USP16 have a better anti-tumor
activity in a tumor model of osteosarcoma and in a 3D system of patient-derived tumor cells.
At the successful completion of this SBIR project we will have identified the best genetic cassettes targeting
USP16 and improving T cell fitness ready for toxicology studies (tumorigenicity and cytokine release syndrome
(CRS) studies).

## Key facts

- **NIH application ID:** 10326136
- **Project number:** 1R43CA265636-01
- **Recipient organization:** DORIAN THERAPEUTICS, INC.
- **Principal Investigator:** Benedetta Nicolis di Robilant
- **Activity code:** R43 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $335,648
- **Award type:** 1
- **Project period:** 2021-09-23 → 2022-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10326136

## Citation

> US National Institutes of Health, RePORTER application 10326136, Epigenetic modulation of cellular aging to improve CAR-T therapy for solid tumors (1R43CA265636-01). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10326136. Licensed CC0.

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