Epigenetic reprogramming to generate novel chondro-osseous stem cells for bone tissue engineering

表观遗传重编程产生用于骨组织工程的新型软骨骨干细胞

基本信息

批准号：
10215392
负责人：
Alan ROBERT Davis
金额：
$ 17.07万
依托单位：
BAYLOR COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-15 至 2022-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10215392
关键词：
ATAC-seq Active Sites Adult Affinity Algorithms Astrocytes Automobile Driving BMP2 gene Bone Diseases Bone Regeneration Bone Tissue Cartilage Cell Maturation Cell physiology Cells Cellular Assay Chondrocytes Chromatin Cluster Analysis CpG dinucleotide DNA Data Data Set Development EGR2 gene Embryo Engineering Enhancers Enterobacteria phage P1 Cre recombinase Epigenetic Process Erythrocytes Exposure to GLAST Protein Generations Genes Genetic Transcription Genomics Glutamate Transporter Grant Heterotopic Ossification Histones Home Hour Label Lead Link Literature Malignant Neoplasms Methyltransferase Modeling Molecular Mus Myelin Basic Proteins N-Cadherin Natural regeneration Nerve Neural Crest Nucleosomes Ontology Osteoblasts Osteogenesis PMP22 gene Pathway interactions Peripheral Nerves Phenotype Phospholipids Physiologic pulse Polycomb Population Process Proteins Red nucleus structure Reporter Reporting Repression Repressor Proteins Role Schwann Cells Site Synapses Tamoxifen Technology Testing Tissue Engineering Tissues Tomatoes Transcript Transposase Variant aortic valve disorder beta Tubulin blastomere structure bone calcification cancer stem cell cell type chromatin remodeling embryo tissue epigenetic memory epigenetic regulation epithelial to mesenchymal transition experimental study fetal reactivity genomic platform histone methylation in vivo in vivo evaluation limb regeneration nerve stem cell neurogranin next generation sequencing novel progenitor promoter relating to nervous system repair model repaired single-cell RNA sequencing soft tissue stem stem cells stem-like cell tissue regeneration transcription factor transcriptome

项目摘要

Recent studies to harness the capacity of BMP2 to induce de novo bone formation for tissue and/or limb regeneration resulted in the identification of a novel chondro-osseous progenitor that is primed to engraft into sites of active bone formation, even when delivered systemically. These cells were identified using lineage tracing for glial high affinity glutamate transporter GLAST, which has been shown to be restricted to neural stem cells, astrocytes, and chondrocytes. Transcriptome analysis, using single cell RNAseq, led to the identification of a highly replicating stem cell that appears to undergo an epithelial to mesenchymal transition (EMT) to become a cell that expresses a large number of chondrocyte and osteoblast associated transcripts similar to a recently reported periosteal stem cell. Surprisingly, while the replicating stem cell expressed GLAST as well as neurogranin, another synaptic protein, and β-tubulin 3/TUJ1; whereas mature chondrocytes did not express this protein supporting its neural association. In further support several or all clusters also expressed several Schwann cell markers including N-cadherin, Krox20, and myelin basic protein. Transcriptome analysis also revealed that this cluster is highly expressing H2A.Z and SET methyltransferase with decreasing synthesis in other clusters/cells correlating with pseudotime or cell maturation. The data collectively has led us to hypothesize that Schwann cells upon exposure to BMP2 undergo epigenetic reprogramming and reactivation of primed tissue specific developmental enhancers leading to a neural crest progenitor/stem cell phenotype. Further, these stem/progenitors are then able to undergo re-differentiation into chondrocytes and osteoblasts. To test this hypothesis, we propose to utilize the phospholipid protein 1 (Plp1) promoter driving Cre recombinase and tomato redfloxSTOPflox mouse to be able to initially label Schwann cells and allow for reporter expression in downstream reprogrammed cells. GLAST-TR+ cells obtained 48 hours after delivery of BMP2, will also be included to obtain potential earlier clusters. GLAST-TR+ and/or Plp1-TR+ cells from soft tissues surrounding the region of de novo bone formation 24 hours after delivery of BMP2 and subjected to single cell RNAseq and single cell ATACseq, which will identify changes in chromatin within the cell populations that might reflect reprogramming of the cells to the highly replicative stem cell. The resultant data will be correlated with the scRNAseq data to provide a molecular mechanism for the generation of these cells. From this data one can then start to envision ex vivo reprogramming of cells to form these chondro- osseous progenitors that will home to sites of active bone formation.

最近的研究，以利用BMP2诱导从头骨形成的能力和/或肢体再生导致鉴定出一种新型的软骨骨祖细胞即使在系统地交付时，也进入活跃的骨形成部位。使用谱系识别这些细胞追踪神经胶质高亲和力谷氨酸转运蛋白胶状，已显示为中性干细胞，星形胶质细胞和软骨细胞。使用单细胞RNASEQ的转录组分析导致鉴定高度复制的干细胞，该干细胞似乎经历了上皮到间充质转变（EMT）成为一个表达大量软骨细胞和成骨细胞相关的成绩单的细胞类似于最近报道的骨膜干细胞。令人惊讶的是，复制的干细胞表达 Glast和Neurogranin，另一种合成蛋白和β-微管蛋白3/Tuj1；而成熟的软骨细胞没有表达这种支持其神经关联的蛋白质。在进一步支持几个或所有集群中表达了几种schwann细胞标记，包括N-钙粘蛋白，KROX20和髓磷脂碱性蛋白。转录组分析还表明，该簇高度表达H2A.Z和固定的甲基转移酶随着其他簇/细胞的合成降低，与假次或细胞成熟相关。数据总体使我们假设暴露于BMP2时Schwann细胞经历表观遗传启动组织特异性发育增强剂的重新编程和重新激活导致神经rest 祖细胞/干细胞表型。此外，这些茎/祖细胞可以重新分化为软骨细胞和成骨细胞。为了检验这一假设，我们建议利用磷脂蛋白1（PLP1）启动子驱动CRE重组酶和番茄Redfloxstopflox小鼠能够最初标记Schwann细胞并允许在下游重编程的细胞中表达报告。 Glast-Tr+细胞获得了48小时交付BMP2后，还将包括以获取潜在的较早集群。 Glast-Tr+和/或PLP1-TR+ BMP2和受到单细胞RNASEQ和单细胞Atacseq的约束，该细胞将确定染色质的变化可能反映细胞重编程为高度复制干细胞的细胞群。结果数据将与SCRNASEQ数据相关，以提供用于生成这些的分子机制细胞。然后，从这些数据中，可以开始设想细胞的离体重编程，以形成这些软骨骨祖细胞会回家，以造成活跃的骨形成部位。

项目成果

期刊论文数量（6）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

MAGI1 inhibits interferon signaling to promote influenza A infection.

DOI：
10.3389/fcvm.2022.791143
发表时间：
2022
期刊：
Frontiers in cardiovascular medicine
影响因子：
3.6
作者：
通讯作者：

A Population of M2 Macrophages Associated With Bone Formation.

DOI：
10.3389/fimmu.2021.686769
发表时间：
2021
期刊：
Frontiers in immunology
影响因子：
7.3
作者：
Olmsted-Davis E;Mejia J;Salisbury E;Gugala Z;Davis AR
通讯作者：
Davis AR

Free Cholesterol Bioavailability and Atherosclerosis.

DOI：
10.1007/s11883-022-01011-z
发表时间：
2022-05
期刊：
CURRENT ATHEROSCLEROSIS REPORTS
影响因子：
5.8
作者：
Abe, Rei J.;Abe, Jun-ichi;Nguyen, Minh T. H.;Olmsted-Davis, Elizabeth A.;Mamun, Abrar;Banerjee, Priyanka;Cooke, John P.;Fang, Longhou;Pownall, Henry;Le, Nhat-Tu
通讯作者：
Le, Nhat-Tu

Cancer treatment-induced NAD+ depletion in premature senescence and late cardiovascular complications.