# Molecular Determinants of Hemogenic Endothelium

> **NIH NIH R01** · UNIVERSITY OF WISCONSIN-MADISON · 2021 · $495,137

## Abstract

ABSTRACT
Progress in cellular reprogramming technologies has created alternative platforms for scalable production of
blood cells for transfusion, immunotherapies and transplantation through inducing pluripotency in somatic cells.
However, even with advances hematopoietic differentiation methods, primitive wave of hematopoiesis
dominates pluripotent stem cell (PSC) differentiation cultures and markers that distinguish primitive and
definitive lymphomyeloid hematopoiesis remains largely unknown. Thus, further translation of hPSCs to
hematology clinic requires a better understanding of the molecular program guiding definitive lymphomyeloid
hematopoiesis. During development, lymphoid progenitors and hematopoietic stem cells (HSCs) arise from
hemogenic endothelium (HE) lining arteries, but not veins. The lack of venous contribution to HSCs along with
the common signaling pathways required for both arterial fate acquisition and HSC development, led to the
hypothesis that arterial specification is a critical prerequisite for HSC formation. However, a direct progenitor-
progeny link between arterial endothelium and definitive lymphomyeloid hematopoiesis has never been
demonstrated. In present application, we propose to prove the hypothesis that arterial specification is an
essential prerequisite for definitive hematopoiesis and demonstrate that promotion of arterial patterning of HE
can provide a novel strategy to aid in generating of lymphoid cells from hPSCs for immunotherapies. In aim 1,
we will identify arterial type of HE (AHE) and demonstrate a direct progenitor-progeny link between AHE and
definitive lymphomyeloid hematopoiesis using arterial-specific enhancer-Cre tracing system. In aim 2, we will
demonstrate that arterial program activation is essential for establishing definitive lymphomyeloid
hematopoietic program. We will show that enhancement of definitive hematopoietic program from hPSCs can
be achieved through activation of arterial program with arteriogenic ETS and SOXF transcription factors (TF),
and modulation of the molecular pathways involved in arteriogenesis using small molecules. In contrast, we will
show that inhibiting arterialization following HE specification abrogates definitive hematopoiesis. Using
RNAseq and ChipSeq analysis we will identify a gene regulatory network connecting arterial and definitive
hematopoietic programs. In aim 3, based on the knowledge gained in understanding the role of arteriogenic
factors in lymphopoiesis, we will develop a forward programming system for T cell generation from hPSCs
using modified mRNA and assess their suitability for CAR-T cell therapies in vivo. Overall, the proposed
studies will establish for the first time a molecular link between arterial programming and definitive
hematopoiesis, and provide evidence that promoting arterial patterning in hPSC cultures can aid to in vitro
approaches to instruct definitive hematopoiesis with lymphoid potentials from hPSCs. In addition, we will off...

## Key facts

- **NIH application ID:** 10187643
- **Project number:** 5R01HL142665-04
- **Recipient organization:** UNIVERSITY OF WISCONSIN-MADISON
- **Principal Investigator:** Igor I. Slukvin
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $495,137
- **Award type:** 5
- **Project period:** 2018-09-01 → 2023-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10187643

## Citation

> US National Institutes of Health, RePORTER application 10187643, Molecular Determinants of Hemogenic Endothelium (5R01HL142665-04). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10187643. Licensed CC0.

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