Cargando…

Single Cell RNA-Seq Analysis of Human Red Cells

Human red blood cells (RBCs), or erythrocytes, are the most abundant blood cells responsible for gas exchange. RBC diseases affect hundreds of millions of people and impose enormous financial and personal burdens. One well-recognized, but poorly understood feature of RBC populations within the same...

Descripción completa

Detalles Bibliográficos
Autores principales: Jain, Vaibhav, Yang, Wen-Hsuan, Wu, Jianli, Roback, John D., Gregory, Simon G., Chi, Jen-Tsan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9065680/
https://www.ncbi.nlm.nih.gov/pubmed/35514346
http://dx.doi.org/10.3389/fphys.2022.828700
_version_ 1784699637779660800
author Jain, Vaibhav
Yang, Wen-Hsuan
Wu, Jianli
Roback, John D.
Gregory, Simon G.
Chi, Jen-Tsan
author_facet Jain, Vaibhav
Yang, Wen-Hsuan
Wu, Jianli
Roback, John D.
Gregory, Simon G.
Chi, Jen-Tsan
author_sort Jain, Vaibhav
collection PubMed
description Human red blood cells (RBCs), or erythrocytes, are the most abundant blood cells responsible for gas exchange. RBC diseases affect hundreds of millions of people and impose enormous financial and personal burdens. One well-recognized, but poorly understood feature of RBC populations within the same individual are their phenotypic heterogeneity. The granular characterization of phenotypic RBC variation in normative and disease states may allow us to identify the genetic determinants of red cell diseases and reveal novel therapeutic approaches for their treatment. Previously, we discovered diverse RNA transcripts in RBCs that has allowed us to dissect the phenotypic heterogeneity and malaria resistance of sickle red cells. However, these analyses failed to capture the heterogeneity found in RBC sub-populations. To overcome this limitation, we have performed single cell RNA-Seq to analyze the transcriptional heterogeneity of RBCs from three adult healthy donors which have been stored in the blood bank conditions and assayed at day 1 and day 15. The expression pattern clearly separated RBCs into seven distinct clusters that include one RBC cluster that expresses HBG2 and a small population of RBCs that express fetal hemoglobin (HbF) that we annotated as F cells. Almost all HBG2-expessing cells also express HBB, suggesting bi-allelic expression in single RBC from the HBG2/HBB loci, and we annotated another cluster as reticulocytes based on canonical gene expression. Additional RBC clusters were also annotated based on the enriched expression of NIX, ACVR2B and HEMGN, previously shown to be involved in erythropoiesis. Finally, we found the storage of RBC was associated with an increase in the ACVR2B and F-cell clusters. Collectively, these data indicate the power of single RBC RNA-Seq to capture and discover known and unexpected heterogeneity of RBC population.
format Online
Article
Text
id pubmed-9065680
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-90656802022-05-04 Single Cell RNA-Seq Analysis of Human Red Cells Jain, Vaibhav Yang, Wen-Hsuan Wu, Jianli Roback, John D. Gregory, Simon G. Chi, Jen-Tsan Front Physiol Physiology Human red blood cells (RBCs), or erythrocytes, are the most abundant blood cells responsible for gas exchange. RBC diseases affect hundreds of millions of people and impose enormous financial and personal burdens. One well-recognized, but poorly understood feature of RBC populations within the same individual are their phenotypic heterogeneity. The granular characterization of phenotypic RBC variation in normative and disease states may allow us to identify the genetic determinants of red cell diseases and reveal novel therapeutic approaches for their treatment. Previously, we discovered diverse RNA transcripts in RBCs that has allowed us to dissect the phenotypic heterogeneity and malaria resistance of sickle red cells. However, these analyses failed to capture the heterogeneity found in RBC sub-populations. To overcome this limitation, we have performed single cell RNA-Seq to analyze the transcriptional heterogeneity of RBCs from three adult healthy donors which have been stored in the blood bank conditions and assayed at day 1 and day 15. The expression pattern clearly separated RBCs into seven distinct clusters that include one RBC cluster that expresses HBG2 and a small population of RBCs that express fetal hemoglobin (HbF) that we annotated as F cells. Almost all HBG2-expessing cells also express HBB, suggesting bi-allelic expression in single RBC from the HBG2/HBB loci, and we annotated another cluster as reticulocytes based on canonical gene expression. Additional RBC clusters were also annotated based on the enriched expression of NIX, ACVR2B and HEMGN, previously shown to be involved in erythropoiesis. Finally, we found the storage of RBC was associated with an increase in the ACVR2B and F-cell clusters. Collectively, these data indicate the power of single RBC RNA-Seq to capture and discover known and unexpected heterogeneity of RBC population. Frontiers Media S.A. 2022-04-20 /pmc/articles/PMC9065680/ /pubmed/35514346 http://dx.doi.org/10.3389/fphys.2022.828700 Text en Copyright © 2022 Jain, Yang, Wu, Roback, Gregory and Chi. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Physiology
Jain, Vaibhav
Yang, Wen-Hsuan
Wu, Jianli
Roback, John D.
Gregory, Simon G.
Chi, Jen-Tsan
Single Cell RNA-Seq Analysis of Human Red Cells
title Single Cell RNA-Seq Analysis of Human Red Cells
title_full Single Cell RNA-Seq Analysis of Human Red Cells
title_fullStr Single Cell RNA-Seq Analysis of Human Red Cells
title_full_unstemmed Single Cell RNA-Seq Analysis of Human Red Cells
title_short Single Cell RNA-Seq Analysis of Human Red Cells
title_sort single cell rna-seq analysis of human red cells
topic Physiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9065680/
https://www.ncbi.nlm.nih.gov/pubmed/35514346
http://dx.doi.org/10.3389/fphys.2022.828700
work_keys_str_mv AT jainvaibhav singlecellrnaseqanalysisofhumanredcells
AT yangwenhsuan singlecellrnaseqanalysisofhumanredcells
AT wujianli singlecellrnaseqanalysisofhumanredcells
AT robackjohnd singlecellrnaseqanalysisofhumanredcells
AT gregorysimong singlecellrnaseqanalysisofhumanredcells
AT chijentsan singlecellrnaseqanalysisofhumanredcells