Project description:We performed scRNA-seq of non-hematopoietic cells from non-treated control (WT), BMT and aGVHD mice. Unsupervised clustering, sub-clustering based on gene expression and trajectory analysis converged on the conclusion that aGVHD likely disrupts MSC differentiation potential, especially osteogenesis.
Project description:BM cells were isolated from C57BL/6 (CD45.2) Tet2–/– mice and Tet2+/+ littermates, as well as from congenic C57BL/6.SJL (CD45.1) mice (all genotypes purchased from JAX). lethally irradiated (9.5 Gy) CD45.1 mice received 10% Tet2–/– CD45.2 BM cells and 90% WT CD45.1 BM cells (2x105 Tet2–/– CD45.2 BM cells and 18x105 WT CD45.1 BM cells; total of 2x106 BM cells/recipient mouse). This group was designated ‘10% Tet2–/– BMT’ (experimental). For the control group that was designated ‘10% Tet2+/+ BMT’, lethally irradiated (9.5 Gy) CD45.1 mice received 10% Tet2+/+ CD45.2 BM cells and 90% WT CD45.1 cells (2x105 Tet2+/+ CD45.2 BM cells & 18x105 WT CD45.1 BM cells, thus exclusively TET2-sufficient cells; total of 2x106 BM cells/recipient mouse). At 12 weeks post-BMT (when there is complete reconstitution of hematopoiesis from transplanted long-term HSCs), ‘10% Tet2–/– BMT’ and ‘10% Tet2+/+ BMT’ mice either received no further treatment (and examined for natural periodontal bone loss) or were subjected to LIP for 5 days.
Project description:Donor OTUD1 deficiency significanltly ameliorates the severity of aGVHD mice. We used single cell RNA sequencing (scRNA-seq) to analyze the immune cells in aGVHD mice transferred wild type or OTUD1-deficient cells.
Project description:We report the application of next generation sequencing technology for high-throughput transcriptom profiling of mucosal associated invariant T (MAIT) cells from patients with or without acute graft versus host disease (aGVHD) post allogenetic hematopoietic stem cell transplantation, and the healthy subjects as the contorls. Totally, MAIT cells from 4 patients with aGVHD, 3 patients without aGVHD and 3 healthy controls were detected in this study. Gene Set Enrichment Analysis (GSEA) on the sequencing data show the enrichments of interferon alpha response pathway in recipients with aGVHD and recipients without aGVHD, in comparison with healthy controls. Enrichment of primary immunodeficiency gene set is found in recipients without aGVHD when compared with aGVHD patients and healthy controls. This study provides some preliminary investigation to the mechanical mechanism of MAIT cells in the aGVHD development.
Project description:The goal of this study are to identify critical biomarkers distinguishing alloHSCT recipients with aGvHD from alloHSCT recipients without aGvHD in two separate cohorts. Total RNA was isolated from the peripheral blood mononucleated cells and then quantified on a Bioanalyzer. Whole transcriptome sequencing libraries (3 pairs of aGvHD and controls, 6 libraries in total) were prepared following the manufacturer’s instructions for the Whole Transcriptome Sample Prep Kit (Illumina) by GENEWIZ Company. 1148 presented significant alterations in cases with aGvHD (t test, FDR corrected P < 0.05, |fold change|>1.5). Transcriptomic pathway analysis showed that the GPL metabolism pathway was one of the most significant pathways with Bonferroni correction P value (Q value) less than 0.001, indicating that it was a critical pathway related to the development of aGvHD.
Project description:RNAs isolated from mouse T cells were processed for sequencing. Naive T cells were wer isoalted from either from WT or Wapl deficiency T cells. The tranfered T cells (either WT or Wapl deficient) were isolated on day 7 after allogeinic or syngeneic BMT. The main purposes of these experiment are to seeking the impact of Wapl KO on gene expression in T cells either in naive status or after BMT.
