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Biomechanical forces promote blood development through prostaglandin E2 and the cAMP-PKA signaling axis.

Diaz MF, Li N, Lee HJ, Adamo L, Evans SM, Willey HE, Arora N, Torisawa YS, Vickers DA, Morris SA, Naveiras O, Murthy SK, Ingber DE, Daley GQ, García-Cardeña G, Wenzel PL - J. Exp. Med. (2015)

Bottom Line: Furthermore, Ncx1 heartbeat mutants, as well as static cultures of AGM, exhibit lower levels of expression of prostaglandin synthases and reduced phosphorylation of the cAMP response element-binding protein (CREB).Similar to flow-exposed cultures, transient treatment of AGM with the synthetic analogue 16,16-dimethyl-PGE2 stimulates more robust engraftment of adult recipients and greater lymphoid reconstitution.These data provide one mechanism by which biomechanical forces induced by blood flow modulate hematopoietic potential.

View Article: PubMed Central - HTML - PubMed

Affiliation: Program in Children's Regenerative Medicine, Department of Pediatric Surgery, Center for Stem Cell and Regenerative Medicine, The Brown Foundation Institute of Molecular Medicine, and Immunology Program, Graduate School of Biomedical Sciences, University of Texas Health Science Center at Houston, Houston, TX 77030 Program in Children's Regenerative Medicine, Department of Pediatric Surgery, Center for Stem Cell and Regenerative Medicine, The Brown Foundation Institute of Molecular Medicine, and Immunology Program, Graduate School of Biomedical Sciences, University of Texas Health Science Center at Houston, Houston, TX 77030 Program in Children's Regenerative Medicine, Department of Pediatric Surgery, Center for Stem Cell and Regenerative Medicine, The Brown Foundation Institute of Molecular Medicine, and Immunology Program, Graduate School of Biomedical Sciences, University of Texas Health Science Center at Houston, Houston, TX 77030.

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WSS induces hematopoietic gene expression and progenitor activity. E9.5 PSp- or E10.5 AGM-derived cells were cultured for 36 h in the presence of 5 dyn/cm2 WSS or static (<0.0001 dyn/cm2) conditions. (A) qRT-PCR of E9.5 PSp demonstrates that WSS induces widespread up-regulation of genes known to be critical regulators of definitive hematopoiesis and lymphopoiesis (n = 5 independent experiments; two-tailed Student’s t test or Mann–Whitney rank sum: *, P < 0.05; **, P < 0.01). (B) Representative flow cytometry plots show distribution of hemogenic endothelial markers (CD144/VE-Cadherin and c-kit) in the cultured populations after 36 h of WSS exposure. Live (DAPI−) CD144+ and CD144+ ckit+ gates are depicted. (C) CD144+ ckit+ CD45−/+ populations are increased by WSS (n = 4 independent experiments; two-tailed Student’s t test: *, P < 0.05). Data are represented as mean ± SEM.
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fig1: WSS induces hematopoietic gene expression and progenitor activity. E9.5 PSp- or E10.5 AGM-derived cells were cultured for 36 h in the presence of 5 dyn/cm2 WSS or static (<0.0001 dyn/cm2) conditions. (A) qRT-PCR of E9.5 PSp demonstrates that WSS induces widespread up-regulation of genes known to be critical regulators of definitive hematopoiesis and lymphopoiesis (n = 5 independent experiments; two-tailed Student’s t test or Mann–Whitney rank sum: *, P < 0.05; **, P < 0.01). (B) Representative flow cytometry plots show distribution of hemogenic endothelial markers (CD144/VE-Cadherin and c-kit) in the cultured populations after 36 h of WSS exposure. Live (DAPI−) CD144+ and CD144+ ckit+ gates are depicted. (C) CD144+ ckit+ CD45−/+ populations are increased by WSS (n = 4 independent experiments; two-tailed Student’s t test: *, P < 0.05). Data are represented as mean ± SEM.

Mentions: Previously, we described specific characteristics of WSS that induce hematopoietic progenitor activity in two-dimensional adherent cultures of dissociated AGM (Adamo et al., 2009). In the present study, we hypothesized that WSS may play a role in specification of HSCs that support life-long adult hematopoiesis and so examined developmental time points that precede definitive HSC emergence in cells derived from the anlage of the AGM known as the PSp at embryonic day (E) 9.5 and from the AGM at E10.5. Runx1 and Myb, two master regulators of hematopoiesis, are transcriptionally up-regulated by shear stress typical of embryonic murine blood flow (5 dyn/cm2), as are other genes required for definitive hematopoiesis and lymphopoiesis, including Bcl11a, Etv6, Gata2, Gata3, Tcf, Rag1, and Rag2 (Fig. 1 A). Analysis of cell surface phenotype after WSS confirmed increases in two markers of hemogenic endothelium, CD144/VE-Cadherin and c-kit, in the live (DAPI−) population (Fig. 1 B). We observed a 5.2 ± 1.2–fold increase in the percentage of CD144+ ckit+ cells, a surface phenotype thought to distinguish a subset of endothelial cells with definitive HSC potential (Fig. 1 C; Eilken et al., 2009; Swiers et al., 2013).


Biomechanical forces promote blood development through prostaglandin E2 and the cAMP-PKA signaling axis.

Diaz MF, Li N, Lee HJ, Adamo L, Evans SM, Willey HE, Arora N, Torisawa YS, Vickers DA, Morris SA, Naveiras O, Murthy SK, Ingber DE, Daley GQ, García-Cardeña G, Wenzel PL - J. Exp. Med. (2015)

WSS induces hematopoietic gene expression and progenitor activity. E9.5 PSp- or E10.5 AGM-derived cells were cultured for 36 h in the presence of 5 dyn/cm2 WSS or static (<0.0001 dyn/cm2) conditions. (A) qRT-PCR of E9.5 PSp demonstrates that WSS induces widespread up-regulation of genes known to be critical regulators of definitive hematopoiesis and lymphopoiesis (n = 5 independent experiments; two-tailed Student’s t test or Mann–Whitney rank sum: *, P < 0.05; **, P < 0.01). (B) Representative flow cytometry plots show distribution of hemogenic endothelial markers (CD144/VE-Cadherin and c-kit) in the cultured populations after 36 h of WSS exposure. Live (DAPI−) CD144+ and CD144+ ckit+ gates are depicted. (C) CD144+ ckit+ CD45−/+ populations are increased by WSS (n = 4 independent experiments; two-tailed Student’s t test: *, P < 0.05). Data are represented as mean ± SEM.
© Copyright Policy - openaccess
Related In: Results  -  Collection

License 1 - License 2
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getmorefigures.php?uid=PMC4419354&req=5

fig1: WSS induces hematopoietic gene expression and progenitor activity. E9.5 PSp- or E10.5 AGM-derived cells were cultured for 36 h in the presence of 5 dyn/cm2 WSS or static (<0.0001 dyn/cm2) conditions. (A) qRT-PCR of E9.5 PSp demonstrates that WSS induces widespread up-regulation of genes known to be critical regulators of definitive hematopoiesis and lymphopoiesis (n = 5 independent experiments; two-tailed Student’s t test or Mann–Whitney rank sum: *, P < 0.05; **, P < 0.01). (B) Representative flow cytometry plots show distribution of hemogenic endothelial markers (CD144/VE-Cadherin and c-kit) in the cultured populations after 36 h of WSS exposure. Live (DAPI−) CD144+ and CD144+ ckit+ gates are depicted. (C) CD144+ ckit+ CD45−/+ populations are increased by WSS (n = 4 independent experiments; two-tailed Student’s t test: *, P < 0.05). Data are represented as mean ± SEM.
Mentions: Previously, we described specific characteristics of WSS that induce hematopoietic progenitor activity in two-dimensional adherent cultures of dissociated AGM (Adamo et al., 2009). In the present study, we hypothesized that WSS may play a role in specification of HSCs that support life-long adult hematopoiesis and so examined developmental time points that precede definitive HSC emergence in cells derived from the anlage of the AGM known as the PSp at embryonic day (E) 9.5 and from the AGM at E10.5. Runx1 and Myb, two master regulators of hematopoiesis, are transcriptionally up-regulated by shear stress typical of embryonic murine blood flow (5 dyn/cm2), as are other genes required for definitive hematopoiesis and lymphopoiesis, including Bcl11a, Etv6, Gata2, Gata3, Tcf, Rag1, and Rag2 (Fig. 1 A). Analysis of cell surface phenotype after WSS confirmed increases in two markers of hemogenic endothelium, CD144/VE-Cadherin and c-kit, in the live (DAPI−) population (Fig. 1 B). We observed a 5.2 ± 1.2–fold increase in the percentage of CD144+ ckit+ cells, a surface phenotype thought to distinguish a subset of endothelial cells with definitive HSC potential (Fig. 1 C; Eilken et al., 2009; Swiers et al., 2013).

Bottom Line: Furthermore, Ncx1 heartbeat mutants, as well as static cultures of AGM, exhibit lower levels of expression of prostaglandin synthases and reduced phosphorylation of the cAMP response element-binding protein (CREB).Similar to flow-exposed cultures, transient treatment of AGM with the synthetic analogue 16,16-dimethyl-PGE2 stimulates more robust engraftment of adult recipients and greater lymphoid reconstitution.These data provide one mechanism by which biomechanical forces induced by blood flow modulate hematopoietic potential.

View Article: PubMed Central - HTML - PubMed

Affiliation: Program in Children's Regenerative Medicine, Department of Pediatric Surgery, Center for Stem Cell and Regenerative Medicine, The Brown Foundation Institute of Molecular Medicine, and Immunology Program, Graduate School of Biomedical Sciences, University of Texas Health Science Center at Houston, Houston, TX 77030 Program in Children's Regenerative Medicine, Department of Pediatric Surgery, Center for Stem Cell and Regenerative Medicine, The Brown Foundation Institute of Molecular Medicine, and Immunology Program, Graduate School of Biomedical Sciences, University of Texas Health Science Center at Houston, Houston, TX 77030 Program in Children's Regenerative Medicine, Department of Pediatric Surgery, Center for Stem Cell and Regenerative Medicine, The Brown Foundation Institute of Molecular Medicine, and Immunology Program, Graduate School of Biomedical Sciences, University of Texas Health Science Center at Houston, Houston, TX 77030.

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Related in: MedlinePlus