PMID- 37843402 OWN - NLM STAT- MEDLINE DCOM- 20240117 LR - 20240117 IS - 2157-6580 (Electronic) IS - 2157-6564 (Print) IS - 2157-6564 (Linking) VI - 13 IP - 1 DP - 2024 Jan 12 TI - T-Cell Mediated Immune Rejection of Beta-2-Microglobulin Knockout Induced Pluripotent Stem Cell-Derived Kidney Organoids. PG - 69-82 LID - 10.1093/stcltm/szad069 [doi] AB - Immune evasive induced pluripotent stem cell (iPSC)-derived kidney organoids, known as "stealth" organoids, hold promise for clinical transplantation. To address immune rejection, we investigated the impact of genetically modifying human leukocyte antigen (HLA) class I in kidney organoids prior to transplantation. By using CRISPR-Cas9, we successfully knocked out beta-2-microglobulin (B2M), resulting in iPSCs devoid of HLA class I surface expression. In vitro, the B2M knockout protected kidney organoids derived from these iPSCs against T-cell rejection. To assess in vivo protection, unmodified (control) and B2M-/- kidney organoids were transplanted into humanized mice engrafted with human peripheral blood mononuclear cells (PBMCs). Successful engraftment of human PBMCs was confirmed, and after 4 weeks, we observed no discernible difference in the infiltration rate, proliferation, or cytotoxicity of CD4+ and CD8+ T cells between control and B2M-/- organoids. Both groups of organoids showed compromised tissue integrity, displaying tubulitis and loss of tubule integrity. Notably, while B2M-/- organoids failed to express HLA class I on their cell surface, there was preexisting expression of HLA class II in both control and B2M-/- organoids transplanted into mice with human PBMCs. HLA class II expression was not limited to antigen-presenting cells but also evident in epithelial cells of the kidney organoid, posing an additional immunological challenge to its transplantation. Consequently, we conclude that B2M knockout alone is insufficient to protect iPSC-derived kidney organoids from T-cell-mediated immune rejection. Additionally, our findings suggest that modulating HLA class II signaling will be necessary to prevent rejection following transplantation. CI - (c) The Author(s) 2023. Published by Oxford University Press. FAU - Gaykema, Lonneke H AU - Gaykema LH AUID- ORCID: 0000-0003-3080-9081 AD - Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine, Leiden University Medical Center (LUMC), Leiden, The Netherlands. AD - Department of Cell and Chemical Biology, Leiden University Medical Center (LUMC), Leiden, The Netherlands. FAU - van Nieuwland, Rianne Y AU - van Nieuwland RY AD - Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine, Leiden University Medical Center (LUMC), Leiden, The Netherlands. FAU - Lievers, Ellen AU - Lievers E AD - Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine, Leiden University Medical Center (LUMC), Leiden, The Netherlands. FAU - Moerkerk, Wessel B J AU - Moerkerk WBJ AD - Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine, Leiden University Medical Center (LUMC), Leiden, The Netherlands. FAU - de Klerk, Juliette A AU - de Klerk JA AD - Department of Cell and Chemical Biology, Leiden University Medical Center (LUMC), Leiden, The Netherlands. FAU - Dumas, Sebastien J AU - Dumas SJ AD - Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine, Leiden University Medical Center (LUMC), Leiden, The Netherlands. AD - The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Leiden University Medical Center (LUMC), Leiden, The Netherlands. FAU - Kers, Jesper AU - Kers J AD - Department of Pathology, Leiden University Medical Center (LUMC), Leiden, The Netherlands. FAU - Zaldumbide, Arnaud AU - Zaldumbide A AD - Department of Cell and Chemical Biology, Leiden University Medical Center (LUMC), Leiden, The Netherlands. FAU - van den Berg, Cathelijne W AU - van den Berg CW AD - Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine, Leiden University Medical Center (LUMC), Leiden, The Netherlands. AD - The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Leiden University Medical Center (LUMC), Leiden, The Netherlands. FAU - Rabelink, Ton J AU - Rabelink TJ AD - Department of Internal Medicine (Nephrology) & Einthoven Laboratory of Vascular and Regenerative Medicine, Leiden University Medical Center (LUMC), Leiden, The Netherlands. AD - The Novo Nordisk Foundation Center for Stem Cell Medicine (reNEW), Leiden University Medical Center (LUMC), Leiden, The Netherlands. LA - eng GR - 2020-01/LUF/Stichting Prof. Jaap de Graeff-Lingling Wiyadharma Fonds/ GR - NNF21CC0073729/Novo Nordisk Foundation/ PT - Journal Article PL - England TA - Stem Cells Transl Med JT - Stem cells translational medicine JID - 101578022 RN - 0 (HLA Antigens) RN - 0 (beta 2-Microglobulin) SB - IM MH - Animals MH - Humans MH - Mice MH - HLA Antigens/metabolism MH - *Induced Pluripotent Stem Cells/metabolism MH - Kidney MH - Leukocytes, Mononuclear MH - Mice, Knockout MH - Organoids MH - beta 2-Microglobulin/metabolism PMC - PMC10785221 OTO - NOTNLM OT - CRISPR OT - T cell OT - differentiation OT - immunogenicity OT - induced pluripotent stem cells (iPSCs) OT - kidney OT - transplantation COIS- None declared. EDAT- 2023/10/16 12:42 MHDA- 2024/01/15 12:42 PMCR- 2023/10/16 CRDT- 2023/10/16 09:43 PHST- 2023/07/20 00:00 [received] PHST- 2023/09/25 00:00 [accepted] PHST- 2024/01/15 12:42 [medline] PHST- 2023/10/16 12:42 [pubmed] PHST- 2023/10/16 09:43 [entrez] PHST- 2023/10/16 00:00 [pmc-release] AID - 7318205 [pii] AID - szad069 [pii] AID - 10.1093/stcltm/szad069 [doi] PST - ppublish SO - Stem Cells Transl Med. 2024 Jan 12;13(1):69-82. doi: 10.1093/stcltm/szad069.