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AGVHD in Italian Children

ABSTRACT: Early signature of aGvHD development in the gut microbiota composition of pediatric patients given allogeneic hematopoietic cell transplantation for hematological disorders

PROVIDER: PRJEB23820 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR2215200_1.fastq.gz	Fastqsanger.gz
	ERR2215200_2.fastq.gz	Fastqsanger.gz
	ERR2215210_1.fastq.gz	Fastqsanger.gz
	ERR2215210_2.fastq.gz	Fastqsanger.gz
	ERR2215220_1.fastq.gz	Fastqsanger.gz

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Transcription profiling of human leukocytes from peripheral blood stem cell transplant patients to investigate acute graft versus host disease aGVHD

Project description:Allogeneic peripheral blood stem cell transplantation (PBSCT) is indicated for the treatment of high-risk hematological malignancies, and in some cases is a cancer cure. A major complication associated with PBSCT (affecting approximately half of recipients) is acute graft-versus-host disease (aGVHD). In aGVHD, alloreactive donor T cells becoming activated by host dendritic cells (DC) in response to pro-inflammatory cytokines released during the conditioning regimen. The result is donor-derived CD4+ T helper-type 1 (Th1) cells maturing and subsequently imposing cytolytic effector responses on host tissues, resulting in multi-organ damage. Despite this well-defined pathophysiological mechanism, there are no definitive markers for predicting aGVHD development or progression to clinically advanced stages. In the current study, we enrolled 8 PBSCT recipients in an IRB-approved study and collected peripheral blood at the time to engraftment. Of these, four patients developed aGVHD within 5-10 days post transplant. The remaining four patients were aGVHD-free. Using total RNA collected from whole blood leukocytes, we analyzed each recipientâs gene expression profile on Affymetrix GeneChip Human Genome U133 Plus 2.0 microarrays. Experiment Overall Design: Samples 1-4 are control patients; 5-8 are aGVHD patients. Peripheral blood was collected at engraftment and used for microarray analysis. In analysing the data, the type of transplant (MUD, related) was used as a covariate to assess if there were significant changes in gene expression between the control (1-4) and aGVHD (5-8) groups, based on a 10% FDR with a pooled error estimate.

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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.

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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.

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