Targeted scRNA-seq analysis of Wnt signaling in bone marrow
Ontology highlight
ABSTRACT: We enriched endothelial cells and other bone marrow cells in both fetal and adult stage to investigate Wnt signaling interaction using targeted scRNA-seq analysis. This analysis facilitate identification of sources of Wnt ligands and detection of Wnt receptor expression in bone marrow. The comparison of fetal and adult stage reveals differences of Wnt signaling in fetal and adult BM.
Project description:Wnt/β-catenin signaling is important in skeletal related-tissue development. We have found that Wnt/β-catenin antagonist XAV could replace traditionally used TGFβ3 as chondrogenic inducer. Here we applied gene expression array to characterize transcriptomic changes between XAV- and TGFβ-induced chondrogenic differentiation in human bone marrow (BM) and induced pluripotent stem cell (iPSC) derived MSCs.
Project description:Multilineage-differentiating stress enduring (Muse) cells are nontumorigenic endogenous pluripotent-like stem cells easily collected from various adult or fetal tissues. The tissue regenerative effects of Muse cells have been demonstrated in many disease models, as they reach damaged sites after intravenous injection to exert pleiotropic effects. Previous reports indicate that several human tissues are readily accessible for Muse cell isolation, including adult tissues such as bone marrow (BM) and embryonic tissues such as Wharton’s Jelly (WJ) from umbilical cord. Wa analyzed the protein repertoires of WJ-Muse and BM-Muse using mass spectrometry-based proteomics.
Project description:Mouse B cell precursors from fetal liver and adult bone marrow generate distinctive B cell progeny when transplanted into immunodeficient recipients, supporting a two-pathway model for B lymphopoiesis, fetal M-bM-^@M-^\B-1M-bM-^@M-^] and adult M-bM-^@M-^\B-2M-bM-^@M-^]. Recently Lin28b was shown to be important for the switch between fetal and adult pathways; however, neither the mechanism of Lin28b action nor the importance of BCR signaling in this process was addressed. Here we report important advances in our understanding of the regulation of B 1/B-2 development. First, modulation of Let-7 in fetal Pro-B cells is sufficient to alter fetal B-1 development to produce B cells resembling the progeny of adult B-2 development. Second, intact BCR signaling is required for generation of B1a B cells from Lin28b-transduced bone marrow progenitors, supporting a requirement for ligand-dependent selection, as is the case for normal B1a B cells. Third, the VH repertore of Lin28b-induced bone marrow B1a B cells differs from that of normal B1a. Finally we identify the Arid3a transcription factor as a key target of Let-7, whose ectopic expression is sufficient to induce B-1 development in adult Pro-B cells and whose knockdown blocks B-1 development in fetal Pro-B cells. 3 individual sorts of Fr.B/C fractions from bone marrow and fetal liver of BALB/c mice
Project description:Genome-wide profiling of RNA expression in two sub-populations of hematopoietic stem cells from mouse Fetal Liver (FL) and adult Bone Marrow (BM)
Project description:Human fetal and adult hematopoietic stem cells (HSC) were obtained from fetal liver, fetal bone marrow (BM), and adult BM. These were injected into human fetal thymic implants in SCID-hu Thy/Liv mice (4-6 separate mice per HSC donor) and allowed to mature into single positive CD4+ (SP4) thymocytes over the course of 7-8 weeks. SP4 thymocytes from injected stem cells were subsequently sort-purified from thymic implants and gene expression was performed.
Project description:Fetal hematopoietic stem and progenitor cells (HSPCs) migrate from fetal liver (FL) to bone marrow (BM) around birth. While adult BM HSPCs and their extrinsic regulation is well studied, little is known about the composition, function, and extrinsic regulation of the first HSPCs to enter the BM. Here, we show that HSPCs colonize multiple fetal bones by E15.5, shift from an MPP2 to an MPP3/4-dominant phenotype by birth, and display little function until E18.5, relative to their FL counterparts. We establish a transcriptional atlas of single perinatal HSPCs, and their putative BM niches, from E15.5 through P0 and show that early fetal BM (FBM) lacks HSPCs with intrinsic stem cell programs and niche cells supportive of HSPCs. In contrast, stem cell programs are preserved in neonatal BM HSPCs, which engage with a niche expressing HSC supportive factors distinct from those seen in adult BM (i.e., IGF).
Project description:The aim of this study is to characterize de genome-wide DNA methylation changes that occur during the fetal-to-adult hemoglobin switch in erythroblasts. We differentiated CD34+ hematopoietic progenitor cells collected for either fetal liver or bone marrow into erythroblasts. After differentiation, cells produce respectively fetal hemoglobin and adult hemoglobin in majority. We compared genome-wide DNA methylation states of fetal-stage erythroblasts to those of adult-stage erythroblasts.