Estimation of cell type proportions from bulk RNA-Seq of porcine whole blood samples using partial reference-free deconvolution

Q4 Biochemistry, Genetics and Molecular Biology Animal Gene Pub Date : 2023-09-01 DOI:10.1016/j.angen.2023.200159
Brittney N. Keel, Amanda K. Lindholm-Perry, Gary A. Rohrer, William T. Oliver
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Abstract

Whole blood has become increasingly utilized in transcriptomic studies because it is easily accessible and can be collected from live animals with minimal invasiveness. However, whole blood represents an extremely complex mixture of cell types, and cell type proportions can confound downstream statistical analyses. Information on cell type proportions may be missing from blood transcriptome studies for a variety of reasons. Experimental approaches for cell counting, such as cell sorting, are arduous and expensive, and therefore may not feasible for studies conducted on a limited budget. Statistical deconvolution can be applied directly to transcriptomic data sets to estimate cell type proportions. In addition to being financially advantageous, computational deconvolution can readily be applied to old datasets, where it may be difficult or impossible to re-analyze for cell type information. In an effort to assist researchers in recovering cell type proportions from porcine whole blood transcriptome samples, we present a manually curated set of porcine blood cell markers that can be utilized in a partial reference-free deconvolution framework to obtain estimates of cell types measured in a standard complete blood count (CBC) panel, which includes neutrophils, lymphocytes, monocytes, eosinophils, basophils, and red blood cells.

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使用部分无参考反褶积从猪全血样品的大量RNA-Seq中估计细胞类型比例
全血在转录组学研究中的应用越来越多,因为它很容易获得,并且可以以最小的入侵性从活体动物身上采集。然而,全血代表了一种极其复杂的细胞类型混合物,细胞类型比例可能会混淆下游的统计分析。由于各种原因,血液转录组研究中可能缺少关于细胞类型比例的信息。细胞计数的实验方法,如细胞分选,既困难又昂贵,因此对于在有限预算下进行的研究来说可能不可行。统计反卷积可以直接应用于转录组数据集,以估计细胞类型比例。除了在经济上有利之外,计算反卷积还可以很容易地应用于旧数据集,在旧数据集中,可能很难或不可能重新分析细胞类型信息。为了帮助研究人员从猪全血转录组样本中恢复细胞类型比例,我们提出了一套手动策划的猪血细胞标志物,可在部分无参考的去卷积框架中使用,以获得在标准全血细胞计数(CBC)面板中测得的细胞类型的估计值,该面板包括中性粒细胞、淋巴细胞、单核细胞,嗜酸性粒细胞、嗜碱性粒细胞和红细胞。
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来源期刊
Animal Gene
Animal Gene Agricultural and Biological Sciences-Insect Science
自引率
0.00%
发文量
16
期刊介绍: Gene Reports publishes papers that focus on the regulation, expression, function and evolution of genes in all biological contexts, including all prokaryotic and eukaryotic organisms, as well as viruses. Gene Reports strives to be a very diverse journal and topics in all fields will be considered for publication. Although not limited to the following, some general topics include: DNA Organization, Replication & Evolution -Focus on genomic DNA (chromosomal organization, comparative genomics, DNA replication, DNA repair, mobile DNA, mitochondrial DNA, chloroplast DNA). Expression & Function - Focus on functional RNAs (microRNAs, tRNAs, rRNAs, mRNA splicing, alternative polyadenylation) Regulation - Focus on processes that mediate gene-read out (epigenetics, chromatin, histone code, transcription, translation, protein degradation). Cell Signaling - Focus on mechanisms that control information flow into the nucleus to control gene expression (kinase and phosphatase pathways controlled by extra-cellular ligands, Wnt, Notch, TGFbeta/BMPs, FGFs, IGFs etc.) Profiling of gene expression and genetic variation - Focus on high throughput approaches (e.g., DeepSeq, ChIP-Seq, Affymetrix microarrays, proteomics) that define gene regulatory circuitry, molecular pathways and protein/protein networks. Genetics - Focus on development in model organisms (e.g., mouse, frog, fruit fly, worm), human genetic variation, population genetics, as well as agricultural and veterinary genetics. Molecular Pathology & Regenerative Medicine - Focus on the deregulation of molecular processes in human diseases and mechanisms supporting regeneration of tissues through pluripotent or multipotent stem cells.
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