# The role of BCL11B in T lineage fate during human thymopoiesis and pluripotent stem cell differentiation

> **NIH NIH R01** · UNIVERSITY OF CALIFORNIA LOS ANGELES · 2024 · $818,795

## Abstract

ABSTRACT/SUMMARY
The functional limitations and logistical challenges of using patient-derived (autologous) products for adoptive T
cell therapy has prompted the exploration of a universal source of “off-the-shelf” T cells generated from self-
renewing PSCs which can be readily genetically engineered to enhance function and expanded without limit.
However current PSC differentiation systems are characterized by low T cell output and concurrent production
of innate lymphoid cells (ILCs). Our preliminary studies suggest that the earliest stages of T cell specification
and commitment seen during PSC differentiation do not fully recapitulate either normal human thymopoiesis or
in vitro models that use definitive hematopoietic stem and progenitor cells (HSPC) to initiate T cell development.
The goal of this proposal is to understand the cellular and molecular differences between normal and
PSC-derived T cell development, with a focus on the role of the transcription factor BCL11B. T cells are
generated in the thymus after notch signaling from the microenvironment triggers a series of transcriptional
events that initiate the T-lineage program in HSPCs; these events first produce early thymic progenitors (ETPs)
(T lineage specification) and then extinguish alternative (non-T) lineage programs in multipotent ETPs (T lineage
commitment). BCL11B is a critical regulator of both of these processes. Our published and preliminary data show
that, in contrast to the mouse model, BCL11B is essential for T cell specification during human thymopoiesis and
initiates the expression of several T-cell genes. Moreover, when BCL11B is overexpressed in cord blood HSPCs,
the T cell program is launched more rapidly and efficiently, even in the absence of notch signaling. Surprisingly
little is known about how the T cell lineage is generated from PSCs. Through scRNA-Seq analysis we have
identified candidate ETPs and their immediate progeny as they emerge from PSC-derived hematopoiesis. We
hypothesize that the rare PSC-derived ETPs in which the T cell program is launched are functionally and
transcriptionally different from ETPs in the thymus, and that these intrinsic differences are detrimental
for the generation of conventional T cells from PSCs. Further, we propose that chromatin remodeling induced
by BCL11B mediates both T lineage specification and the fate decisions between the conventional T cell and
innate lymphoid pathways. Specifically we will: 1. Define the earliest T lineage progenitors generated during PSC
differentiation; 2. Determine the epigenetic underpinnings of T-cell specification in PSC-ATOs and in primary
thymopoiesis; and 3. Define how BCL11B affects conventional T and innate lineage fate choices. These studies
will yield new mechanistic insights about T-cell differentiation that are critical for the development of PSC-derived
T-cell immunotherapies.

## Key facts

- **NIH application ID:** 10877783
- **Project number:** 5R01AI176249-02
- **Recipient organization:** UNIVERSITY OF CALIFORNIA LOS ANGELES
- **Principal Investigator:** Gay M Crooks
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $818,795
- **Award type:** 5
- **Project period:** 2023-07-01 → 2028-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10877783

## Citation

> US National Institutes of Health, RePORTER application 10877783, The role of BCL11B in T lineage fate during human thymopoiesis and pluripotent stem cell differentiation (5R01AI176249-02). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10877783. Licensed CC0.

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