# Stem cell immunoengineering for universal cardiac therapy via CRISPR-Cas9

> **NIH NIH R21** · PENNSYLVANIA STATE UNIVERSITY, THE · 2020 · $214,663

## Abstract

Project Summary:
Human cardiac cells derived from human pluripotent stem cells (hPSCs) hold the
potential to dramatically change the treatment of cardiac diseases and provide new
strategies for rebuilding the adult heart after injury. In order to realize the therapeutic
application of these cells, however, we need to better understand the mechanisms
underlying the immune rejection during allogeneic cell transplantation. Here, the
investigators propose a novel paradigm for control of hPSC immunogenicity, using
recent advances in genome editing (CRISPR-Cas9) to program universal immune
protective hPSC clones. These universal donor stem cells can then differentiate into
human ventricular progenitors (HVPs) for allogeneic transplantation.
The investigator recently developed a new efficient HVP differentiation method and
performed extensive in vivo studies of HVPs and showed that the HVPs are better than
cardiomyocytes in terms of survival, proliferation and generation of vascularized tissue.
In this proposal, the investigator will employ recent advances in understanding the
interaction between the human immune cells and transplanted donor cells and the
CRISPR-Cas9 technique to design universal donor stem cells that confer immune
protective properties for allogeneic transplantation. The investigator will use Cas9 to
engineer stem cells for avoiding T-cell rejection via a gene knockout of beta2-
microglobulin, which is essential for forming functional HLA-I molecules on the cell
surface. In addition, a safe-harbor knockin for overexpression of PDL1 and CTLA4-Ig
genes, which play critical T-cell immune modulation roles in allotransplantation, is also
considered. Furthermore, to avoid the NK-mediated lysis of donor cells, the investigator
will express HLA-E-B2M and HLA-G-B2M to inhibit the CD94/NKG2A and 2DL4 NK cell
inhibitory receptors respectively, which will suppress the NK-mediated lysis of the donor
cells. Last but not least, the investigator will evaluate the performance of engineered
HVPs using in vitro and in vivo models. The investigator will examine the growth of
luciferase-expressing engineered HVPs and control HVPs transplanted into
immunodeficient mice injected with human T cells or NK cells.

## Key facts

- **NIH application ID:** 9880434
- **Project number:** 5R21EB026035-03
- **Recipient organization:** PENNSYLVANIA STATE UNIVERSITY, THE
- **Principal Investigator:** Xiaojun Lian
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $214,663
- **Award type:** 5
- **Project period:** 2018-06-01 → 2022-02-28

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/9880434

## Citation

> US National Institutes of Health, RePORTER application 9880434, Stem cell immunoengineering for universal cardiac therapy via CRISPR-Cas9 (5R21EB026035-03). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/9880434. Licensed CC0.

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