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1.
Dev Cell ; 53(5): 503-513.e5, 2020 06 08.
Article in English | MEDLINE | ID: mdl-32413329

ABSTRACT

Bone marrow (BM) mesenchymal stem and progenitor cells (MSPCs) are a critical constituent of the hematopoietic stem cell (HSC) niche. Previous studies have suggested that the zinc-finger epithelial-mesenchymal transition transcription factor Snai2 (also known as Slug) regulated HSCs autonomously. Here, we show that Snai2 expression in the BM is restricted to the BM stromal compartment where it regulates the HSC niche. Germline or MSPC-selective Snai2 deletion reduces the functional MSPC pool and their mesenchymal lineage output and impairs HSC niche function during homeostasis and after stress. RNA sequencing analysis revealed that Spp1 (osteopontin) expression is markedly upregulated in Snai2-deficient MSPCs. Genetic deletion of Spp1 in Snai2-deficient mice rescues MSPCs' functions. Thus, SNAI2 is a critical regulator of the transcriptional network maintaining MSPCs by the suppression of osteopontin expression.


Subject(s)
Bone Marrow Cells/metabolism , Osteopontin/genetics , Snail Family Transcription Factors/metabolism , Stem Cell Niche , Animals , Bone Marrow Cells/cytology , Cells, Cultured , Gene Deletion , Mice , Mice, Inbred C57BL , Osteopontin/metabolism , Snail Family Transcription Factors/genetics
2.
Sci Rep ; 9(1): 10508, 2019 07 19.
Article in English | MEDLINE | ID: mdl-31324840

ABSTRACT

Aneuploidy has been reported to occur at remarkably high levels in normal somatic tissues using Fluorescence In Situ Hybridization (FISH). Recently, these reports were contradicted by single-cell low-coverage whole genome sequencing (scL-WGS) analyses, which showed aneuploidy frequencies at least an order of magnitude lower. To explain these seemingly contradictory findings, we used both techniques to analyze artificially generated mock aneuploid cells and cells with natural random aneuploidy. Our data indicate that while FISH tended to over-report aneuploidies, a modified 2-probe approach can accurately detect low levels of aneuploidy. Further, scL-WGS tends to underestimate aneuploidy levels, especially in a polyploid background.


Subject(s)
Aneuploidy , In Situ Hybridization, Fluorescence/methods , Single-Cell Analysis , Whole Genome Sequencing/methods , Animals , Cerebral Cortex/cytology , Fibroblasts/ultrastructure , Hepatocytes/ultrastructure , Humans , Interphase , Karyotyping/methods , Mice , Neurons/ultrastructure , Polyploidy , Reproducibility of Results , Sensitivity and Specificity
3.
Carcinogenesis ; 40(8): 937-946, 2019 08 22.
Article in English | MEDLINE | ID: mdl-31169292

ABSTRACT

Sporadic colon cancer accounts for approximately 80% of colorectal cancer (CRC) with high incidence in Western societies strongly linked to long-term dietary patterns. A unique mouse model for sporadic CRC results from feeding a purified rodent Western-style diet (NWD1) recapitulating intake for the mouse of common nutrient risk factors each at its level consumed in higher risk Western populations. This causes sporadic large and small intestinal tumors in wild-type mice at an incidence and frequency similar to that in humans. NWD1 perturbs intestinal cell maturation and Wnt signaling throughout villi and colonic crypts and decreases mouse Lgr5hi intestinal stem cell contribution to homeostasis and tumor development. Here we establish that NWD1 transcriptionally reprograms Lgr5hi cells, and that nutrients are interactive in reprogramming. Furthermore, the DNA mismatch repair pathway is elevated in Lgr5hi cells by lower vitamin D3 and/or calcium in NWD1, paralleled by reduced accumulation of relevant somatic mutations detected by single-cell exome sequencing. In compensation, NWD1 also reprograms Bmi1+ cells to function and persist as stem-like cells in mucosal homeostasis and tumor development. The data establish the key role of the nutrient environment in defining the contribution of two different stem cell populations to both mucosal homeostasis and tumorigenesis. This raises important questions regarding impact of variable human diets on which and how stem cell populations function in the human mucosa and give rise to tumors. Moreover, major differences reported in turnover of human and mouse crypt base stem cells may be linked to their very different nutrient exposures.


Subject(s)
Carcinogenesis/genetics , Colonic Neoplasms/genetics , Stem Cells/metabolism , Animals , Calcium/metabolism , Cell Differentiation/genetics , Cell Proliferation/genetics , Cholecalciferol/metabolism , Colonic Neoplasms/metabolism , Colonic Neoplasms/pathology , Diet, Western/adverse effects , Disease Models, Animal , Homeostasis/genetics , Humans , Intestinal Mucosa/metabolism , Intestines/growth & development , Mice , Nutrition Assessment , Receptors, G-Protein-Coupled/genetics , Signal Transduction/genetics , Wnt Signaling Pathway/genetics
4.
BMC Bioinformatics ; 20(1): 336, 2019 Jun 17.
Article in English | MEDLINE | ID: mdl-31208319

ABSTRACT

BACKGROUND: Numerical chromosomal variation is a hallmark of populations of malignant cells. Identifying the factors that promote numerical chromosomal variation is important for understanding mechanisms of carcinogenesis. However, the ability to quantify and visualize differences in chromosome number between experimentally-defined groups (e.g. control vs treated) obtained from single-cell experiments is currently limited by the lack of user-friendly software. RESULTS: Aneuvis is a web application that allows users to determine whether numerical chromosomal variation exists between experimental treatment groups. The web interface allows users to upload molecular cytogenetic or processed single cell whole-genome sequencing data in a cell-by-chromosome matrix format and automatically generates visualizations and summary statistics that reflect the degree of numeric chromosomal variability. CONCLUSIONS: Aneuvis is the first user-friendly web application to help researchers identify the genetic and environmental perturbations that promote numerical chromosomal variation. Aneuvis is freely available as a web application at https://dpique.shinyapps.io/aneuvis/ and the source code for the application is available at https://github.com/dpique/aneuvis .


Subject(s)
Chromosomes/genetics , Internet , Single-Cell Analysis , Software , Cell Line , DNA Copy Number Variations/genetics , Humans , User-Computer Interface , Whole Genome Sequencing
5.
Alzheimers Dement (N Y) ; 5: 906-917, 2019.
Article in English | MEDLINE | ID: mdl-31890855

ABSTRACT

INTRODUCTION: Women are at increased risk for Alzheimer's disease (AD), but the reason why remains unknown. One hypothesis is that low estrogen levels at menopause increases vulnerability to AD, but this remains unproven. METHODS: We compared neuronal genes upregulated by estrogen in ovariectomized female rhesus macaques with a database of >17,000 diverse gene sets and applied a rare variant burden test to exome sequencing data from 1208 female AD patients with the age of onset < 75 years and 2162 female AD controls. RESULTS: We found a striking overlap between genes upregulated by estrogen in macaques and genes downregulated in the human postmortem AD brain, and we found that estrogen upregulates the APOE gene and that progesterone acts antagonistically to estrogen genome-wide. We also found that female patients with AD have excess rare mutations in the early menopause gene MCM8. DISCUSSION: We show with genomic data that the menopausal loss of estrogen could underlie the increased risk for AD in women.

6.
Nat Commun ; 9(1): 2449, 2018 06 22.
Article in English | MEDLINE | ID: mdl-29934585

ABSTRACT

Endothelial cells (ECs) contribute to haematopoietic stem cell (HSC) maintenance in bone marrow, but the differential contributions of EC subtypes remain unknown, owing to the lack of methods to separate with high purity arterial endothelial cells (AECs) from sinusoidal endothelial cells (SECs). Here we show that the combination of podoplanin (PDPN) and Sca-1 expression distinguishes AECs (CD45- Ter119- Sca-1bright PDPN-) from SECs (CD45- Ter119- Sca-1dim PDPN+). PDPN can be substituted for antibodies against the adhesion molecules ICAM1 or E-selectin. Unexpectedly, prospective isolation reveals that AECs secrete nearly all detectable EC-derived stem cell factors (SCF). Genetic deletion of Scf in AECs, but not SECs, significantly reduced functional HSCs. Lineage-tracing analyses suggest that AECs and SECs self-regenerate independently after severe genotoxic insults, indicating the persistence of, and recovery from, radio-resistant pre-specified EC precursors. AEC-derived SCF also promotes HSC recovery after myeloablation. These results thus uncover heterogeneity in the contribution of ECs in stem cell niches.


Subject(s)
Bone Marrow Cells/metabolism , Endothelial Cells/metabolism , Stem Cell Factor/metabolism , Animals , Antigens, Ly/metabolism , Arteries/cytology , Bone Marrow/blood supply , Bone Marrow Transplantation , Capillaries/cytology , Cell Differentiation/physiology , Cells, Cultured , Female , Hematopoietic Stem Cells/physiology , Male , Membrane Glycoproteins/metabolism , Membrane Proteins/metabolism , Mice , Mice, Inbred C57BL , Mice, Transgenic , Stem Cell Factor/genetics , Stem Cell Niche/physiology , Transplantation Chimera
7.
Science ; 354(6316): 1156-1160, 2016 12 02.
Article in English | MEDLINE | ID: mdl-27738012

ABSTRACT

A single hematopoietic stem cell (HSC) is capable of reconstituting hematopoiesis and maintaining homeostasis by balancing self-renewal and cell differentiation. The mechanisms of HSC division balance, however, are not yet defined. Here we demonstrate, by characterizing at the single-cell level a purified and minimally heterogeneous murine Tie2+ HSC population, that these top hierarchical HSCs preferentially undergo symmetric divisions. The induction of mitophagy, a quality control process in mitochondria, plays an essential role in self-renewing expansion of Tie2+ HSCs. Activation of the PPAR (peroxisome proliferator-activated receptor)-fatty acid oxidation pathway promotes expansion of Tie2+ HSCs through enhanced Parkin recruitment in mitochondria. These metabolic pathways are conserved in human TIE2+ HSCs. Our data thus identify mitophagy as a key mechanism of HSC expansion and suggest potential methods of cell-fate manipulation through metabolic pathways.


Subject(s)
Cell Self Renewal , Hematopoiesis/physiology , Hematopoietic Stem Cells/physiology , Mitochondria/physiology , Mitophagy/physiology , Animals , Cell Separation , Fatty Acids/metabolism , Green Fluorescent Proteins/analysis , Green Fluorescent Proteins/metabolism , Hematopoietic Stem Cells/chemistry , Metabolic Networks and Pathways , Mice , Mice, Inbred C57BL , Mitophagy/genetics , Oxidation-Reduction , Peroxisome Proliferator-Activated Receptors/metabolism , Receptor, TIE-2/analysis , Single-Cell Analysis , Ubiquitin-Protein Ligases/genetics , Ubiquitin-Protein Ligases/metabolism
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