Maintenance of Hypoimmunogenic Features via Regulation of Endogenous Antigen Processing and Presentation Machinery

Ju-Hyun An; Hyebin Koh; Yujin Ahn; Jieun Kim; A-Reum Han; Ji Yoon Lee; Sun-Uk Kim; Jong-Hee Lee

doi:10.3389/fbioe.2022.936584

Maintenance of Hypoimmunogenic Features via Regulation of Endogenous Antigen Processing and Presentation Machinery

Front Bioeng Biotechnol. 2022 Jul 22:10:936584. doi: 10.3389/fbioe.2022.936584. eCollection 2022.

Authors

Ju-Hyun An^{1

2}, Hyebin Koh^{1

2}, Yujin Ahn^{1

2}, Jieun Kim^{3

4}, A-Reum Han⁵, Ji Yoon Lee⁵, Sun-Uk Kim^{1

2}, Jong-Hee Lee^{2

4}

Affiliations

¹ Futuristic Animal Resource and Research Center (FARRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB), Ochang, South Korea.
² Department of Functional Genomics, KRIBB School of Bioscience, Korea University of Science and Technology, Daejeon, South Korea.
³ Department of Medical Life Sciences, College of Medicine, The Catholic University of Korea, Seoul, South Korea.
⁴ National Primate Research Center (NPRC), Korea Research Institute of Bioscience and Biotechnology (KRIBB), Ochang, South Korea.
⁵ CHA Advanced Research Institute, Bundang CHA Hospital, CHA University, Seongnam, South Korea.

Abstract

Universally acceptable donor cells have been developed to address the unmet need for immunotypically matched materials for regenerative medicine. Since forced expression of hypoimmunogenic genes represses the immune response, we established universal pluripotent stem cells (PSCs) by replacing endogenous β2-microglobulin (β2m) with β2m directly conjugated to human leukocyte antigen (HLA)-G, thereby simultaneously suppressing HLA-I expression and the natural killer (NK) cell-mediated immune response. These modified human PSCs retained their pluripotency and differentiation capacity; however, surface presentation of HLA-G was absent from subsequently differentiated cells, particularly cells of neural lineages, due to the downregulation of antigen processing and presentation machinery (APM) genes. Induction of APM genes by overexpression of NLR-family CARD domain-containing 5 (NLRC5) or activator subunit of nuclear factor kappa B (NF-κB) heterodimer (RelA) recovered the surface expression of HLA-G and the hypoimmunogenicity of neural cells. Our findings enhance the utility of hypoimmunogenic cells as universal donors and will contribute to the development of off-the-shelf stem-cell therapeutics.

Keywords: antigen presentation and processing machinery; cell therapy; hypoimmunogenic cell; regenerative cell therapy; universal donor cell.