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Single Cell RNA Sequencing Analysis of 3 Human Living-Donor Liver Transplant Allograft Biopsies after Infusion of Donor-Derived Regulatory Dendritic Cells

L. M. Tran1, T. Tabib2, A. F. Zahorchak1, R. Lafyatis2, M. A. Styn1, A. Humar1, F. G. Lakkis1, D. M. Metes1, A. W. Thomson1

1Department of Surgery, University of Pittsburgh School of Medicine, Pittsburgh, PA, 2Department of Medicine and Immunology, University of Pittsburgh School of Medicine, Pittsburgh, PA

Meeting: 2020 American Transplant Congress

Abstract number: B-382

Keywords: Liver transplantation, Tolerance

Session Information

Date: Saturday, May 30, 2020

Session Name: Poster Session B: Tolerance / Immune Deviation

Session Time: 3:15pm-4:00pm

 Presentation Time: 3:30pm-4:00pm

Location: Virtual

Related Abstracts
  • Fate of Donor-Derived Regulatory Dendritic Cells (DCreg) and Augmentation of Host Regulatory T Cells Following Their Infusion Into Prospective Living-Donor Liver Transplant Recipients
  • Regulatory Dendritic Cell (DCreg) Cell Infusion in Living Donor Liver Transplantation

*Purpose: Single-cell RNA sequencing is an emerging, powerful genomic technology that can describe heterogenous cellular populations of whole solid organs with high resolution. We report on preliminary single-cell RNA sequencing analysis of liver biopsy tissue obtained at time of transplant from three enrolled living-donor liver transplant (LDLTx) patients infused with donor-derived regulatory dendritic (DCreg) cells as part of an ongoing in-human phase I/II clinical trial (NCT03164265).

*Methods: DCregs generated from prospective living donor elutriated monocytes were infused into three LDLTx recipients enrolled in our study 7 days prior to transplantation. An 18-gauge core needle biopsy of the donor hepatic allograft was obtained 1-hour after reperfusion in the recipient and sent for tissue digestion and processing. A subsequent single cell genomics library was prepared using the 10x Genomics platform. Data was normalized and clustered to identify distinct resident cell populations in the transplanted livers after DCreg infusion.

*Results: We generated 6,440 single cell transcriptomes of transplant hepatic tissue from three trial LDLTx recipients and performed unsupervised clustering analysis of gene expression. We uniquely identify 4 clusters of granulocytes/neutrophils and 4 clusters of NK cells in our transcriptional profile that has not been previously reported in single cell analysis on normal human liver. We also demonstrate the presence of a CD16+ monocyte population that represents a distinct non-classical subset of monocytes. Regulatory gene expression profiles across several innate and adaptive immune cell types were represented, including CTLA4 which may indicate the presence of a regulatory T cell population.

*Conclusions: A large number of immune cell subtypes are represented in the immediate post-transplant reperfused liver, distinct from that previously reported in normal human liver. Regulatory gene expression profiles are present and may indicate a heterogenous immune response to DCreg infusion, liver transplantation, or both. This preliminary study provides a proof of concept for the role of next-generation sequencing in defining changes to tissue immune cell landscape as response to stimuli.

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To cite this abstract in AMA style:

Tran LM, Tabib T, Zahorchak AF, Lafyatis R, Styn MA, Humar A, Lakkis FG, Metes DM, Thomson AW. Single Cell RNA Sequencing Analysis of 3 Human Living-Donor Liver Transplant Allograft Biopsies after Infusion of Donor-Derived Regulatory Dendritic Cells [abstract]. Am J Transplant. 2020; 20 (suppl 3). https://atcmeetingabstracts.com/abstract/single-cell-rna-sequencing-analysis-of-3-human-living-donor-liver-transplant-allograft-biopsies-after-infusion-of-donor-derived-regulatory-dendritic-cells/. Accessed March 4, 2021.

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