Possible cooption of a VEGF-driven tubulogenesis program for biomineralization in echinoderms

Biomineralization is the process by which living organisms use minerals to form hard structures that protect and support them. Biomineralization is believed to have evolved rapidly and independently in different phyla utilizing preexisting components. The mechanistic understanding of the regulatory networks that drive biomineralization and their evolution is far from clear. Sea urchin skeletogenesis is an excellent model system for studying both gene regulation and mineral uptake and deposition. The sea urchin calcite spicules are formed within a tubular cavity generated by the skeletogenic cells controlled by vascular endothelial growth factor (VEGF) signaling. The VEGF pathway is essential for biomineralization in echinoderms, while in many other phyla, across metazoans, it controls tubulogenesis and vascularization. Despite the critical role of VEGF signaling in sea urchin spiculogenesis, the downstream program it activates was largely unknown. Here we study the cellular and molecular machinery activated by the VEGF pathway during sea urchin spiculogenesis and reveal multiple parallels to the regulation of vertebrate vascularization. Human VEGF rescues sea urchin VEGF knockdown, vesicle deposition into an internal cavity plays a significant role in both systems, and sea urchin VEGF signaling activates hundreds of genes, including biomineralization and interestingly, vascularization genes. Moreover, five upstream transcription factors and three signaling genes that drive spiculogenesis are homologous to vertebrate factors that control vascularization. Overall, our findings suggest that sea urchin spiculogenesis and vertebrate vascularization diverged from a common ancestral tubulogenesis program, broadly adapted for vascularization and specifically coopted for biomineralization in the echinoderm phylum.

Download Full-text

Possible co-option of a VEGF-driven tubulogenesis program for biomineralization in echinoderms

10.1101/554683 ◽

2019 ◽

Author(s):

Miri Morgulis ◽

Tsvia Gildor ◽

Modi Roopin ◽

Noa Sher ◽

Assaf Malik ◽

...

Keyword(s):

Vascular Endothelial Growth Factor ◽

Growth Factor ◽

Sea Urchin ◽

Regulatory Networks ◽

Critical Role ◽

Vascular Endothelial ◽

Upstream Gene ◽

Vegf Signaling ◽

Vegf Pathway ◽

Endothelial Growth Factor

AbstractBiomineralization is the process in which living organisms use minerals to form hard structures that protect and support them. Biomineralization is believed to have evolved rapidly and independently in different phyla utilizing existing components used for other purposes. The mechanistic understanding of the regulatory networks that drive biomineralization and their evolution is far from clear. The sea urchin skeletogenesis is an excellent model system for studying both gene regulation and mineral uptake and deposition. The sea urchin calcite spicules are formed within a tubular cavity generated by the skeletogenic cells under the control the vascular endothelial growth factor (VEGF) signaling. The VEGF pathway controls tubulogenesis and vascularization across metazoans while its regulation of biomineralization was only observed in echinoderms. Despite the critical role of VEGF signaling in sea urchin spiculogenesis, the downstream program it activates was largely unknown. Here we study the cellular and molecular machinery activated by the VEGF pathway during sea urchin spiculogenesis and reveal multiple parallels to the regulation of tubulogenesis during vertebrate vascularization. Human VEGF rescues sea urchin VEGF knock-down; VEGF-dependent vesicle deposition plays a significant role in both systems and sea urchin VEGF signaling activates hundreds of genes including biomineralization and vascularization genes. Five upstream transcription factors and three signaling genes active in spiculogenesis are homologous to vertebrate factors that regulate vascularization. Overall, our findings suggest that sea urchin spiculogenesis and vertebrate vascularization diverged from a common ancestral tubulogenesis program, broadly adapted for vascularization and specifically co-opted for biomineralization in the echinoderm phylum.Significance statementThe sea urchin calcite spicules and vertebrate blood vessels are quite distinct in their function, yet both have a tubular structure and are controlled by the vascular endothelial growth factor (VEGF) pathway. Here we study the downstream program by which VEGF pathway drives sea urchin spiculogenesis and find remarkable similarities to the control of vertebrate vascularization. The similarities are observed both in the upstream gene regulatory network, in the downstream effector genes and the cellular processes that VEGF signaling controls at the site of the calcite spicule formation. We speculate that sea urchin spiculogenesis and vertebrate vascularization diverged from a common ancestral tubulogenesis program that was co-opted for biomineralization in the echinoderm phylum.

Download Full-text

Endothelial Semaphorin 3fb regulates Vegf pathway-mediated angiogenic sprouting

10.1101/2021.08.03.454978 ◽

2021 ◽

Author(s):

Charlene Watterston ◽

Rami Halabi ◽

Sarah McFarlane ◽

Sarah J Childs

Keyword(s):

Growth Factor ◽

Critical Role ◽

Developmental Time ◽

Vegf Receptor ◽

Growth Factor Signaling ◽

Actin Assembly ◽

Vegf Signaling ◽

Vessel Growth ◽

Vegf Pathway ◽

Endothelial Growth Factor

Vessel growth integrates diverse extrinsic signals with intrinsic signaling cascades to coordinate cell migration and sprouting morphogenesis. The pro-angiogenic effects of Vascular Endothelial Growth Factor (VEGF) are carefully controlled during sprouting to generate an efficiently patterned vascular network. We identify crosstalk between VEGF signaling and that of the secreted ligand Semaphorin 3fb (Sema3fb), one of two zebrafish paralogs of mammalian Sema3F. The sema3fb gene is expressed by endothelial cells in actively sprouting vessels. Loss of sema3fb results in abnormally wide and stunted intersegmental vessel artery sprouts. Although the sprouts initiate at the correct developmental time, they have a reduced migration speed. These sprouts have persistent filopodia and abnormally spaced nuclei suggesting dysregulated control of actin assembly. sema3fb mutants show simultaneously higher expression of pro-angiogenic (VEGF receptor 2 (vegfr2) and delta-like 4 (dll4)) and anti-angiogenic (soluble VEGF receptor 1 (svegfr1)/ soluble Fms Related Receptor Tyrosine Kinase 1 (sflt1)) pathway components. We show increased phospho-ERK staining in migrating angioblasts, consistent with enhanced Vegf activity. Reducing Vegfr2 kinase activity in sema3fb mutants rescues angiogenic sprouting. Our data suggest that Sema3fb plays a critical role in promoting endothelial sprouting through modulating the VEGF signaling pathway, acting as an autocrine cue that modulates intrinsic growth factor signaling.

Download Full-text

Endothelial Semaphorin 3fb regulates Vegf pathway-mediated angiogenic sprouting

PLoS Genetics ◽

10.1371/journal.pgen.1009769 ◽

2021 ◽

Vol 17 (8) ◽

pp. e1009769

Author(s):

Charlene Watterston ◽

Rami Halabi ◽

Sarah McFarlane ◽

Sarah J. Childs

Keyword(s):

Growth Factor ◽

Critical Role ◽

Developmental Time ◽

Vegf Receptor ◽

Growth Factor Signaling ◽

Actin Assembly ◽

Vegf Signaling ◽

Vessel Growth ◽

Vegf Pathway ◽

Endothelial Growth Factor

Download Full-text

Scl isoforms act downstream of etsrp to specify angioblasts and definitive hematopoietic stem cells

Blood ◽

10.1182/blood-2009-09-244640 ◽

2010 ◽

Vol 115 (26) ◽

pp. 5338-5346 ◽

Cited By ~ 42

Author(s):

Xi Ren ◽

Gustavo A. Gomez ◽

Bo Zhang ◽

Shuo Lin

Keyword(s):

Stem Cells ◽

Hematopoietic Stem Cells ◽

Critical Role ◽

Vascular Endothelial ◽

Hematopoietic Stem ◽

Vegf Signaling ◽

Endothelial Growth Factor ◽

Endothelial Development ◽

Definitive Hematopoiesis ◽

Runx1 Expression

Abstract Recent lineage studies suggest that hematopoietic stem cells (HSCs) may be derived from endothelial cells. However, the genetic hierarchy governing the emergence of HSCs remains elusive. We report here that zebrafish ets1-related protein (etsrp), which is essential for vascular endothelial development, also plays a critical role in the initiation of definitive hematopoiesis by controlling the expression of 2 stem cell leukemia (scl) isoforms (scl-α and scl-β) in angioblasts. In etsrp morphants, which are deficient in endothelial and HSC development, scl-α alone partially rescues angioblast specification, arterial-venous differentiation, and the expression of HSC markers, runx1 and c-myb, whereas scl-β requires angioblast rescue by fli1a to restore runx1 expression. Interestingly, when vascular endothelial growth factor (Vegf) signaling is inhibited, HSC marker expression can still be restored by scl-α in etsrp morphants, whereas the rescue of arterial ephrinb2a expression is blocked. Furthermore, both scl isoforms partially rescue runx1 but not ephrinb2a expression in embryos deficient in Vegf signaling. Our data suggest that downstream of etsrp, scl-α and fli1a specify the angioblasts, whereas scl-β further initiates HSC specification from this angioblast population, and that Vegf signaling acts upstream of scl-β during definitive hematopoiesis.

Download Full-text

Expression of Tumor-Derived Vascular Endothelial Growth Factor and Its Receptors Is Associated With Outcome in Early Squamous Cell Carcinoma of the Lung

Journal of Clinical Oncology ◽

10.1200/jco.2011.37.4231 ◽

2012 ◽

Vol 30 (10) ◽

pp. 1129-1136 ◽

Cited By ~ 47

Author(s):

María J. Pajares ◽

Jackeline Agorreta ◽

Marta Larrayoz ◽

Aurélien Vesin ◽

Teresa Ezponda ◽

...

Keyword(s):

Lung Cancer ◽

Vascular Endothelial Growth Factor ◽

Squamous Cell Carcinoma ◽

Growth Factor ◽

Vascular Endothelial ◽

Vegf Signaling ◽

Vegf Pathway ◽

Cell Expression ◽

Endothelial Growth Factor ◽

Histologic Subtypes

PurposeAntiangiogenic therapies targeting the vascular endothelial growth factor (VEGF) pathway have yielded more modest clinical benefit to patients with non–small-cell lung cancer (NSCLC) than initially expected. Clinical data suggest a distinct biologic role of the VEGF pathway in the different histologic subtypes of lung cancer. To clarify the influence of histologic differentiation in the prognostic relevance of VEGF-mediated signaling in NSCLC, we performed a concomitant analysis of the expression of three key elements of the VEGF pathway in the earliest stages of the following two principal histologic subtypes: squamous cell carcinoma (SCC) and adenocarcinoma (ADC).Patients and MethodsWe evaluated tumor cell expression of VEGF, VEGF receptor (VEGFR) 1, and VEGFR2 using automatic immunostaining in a series of 298 patients with early-stage NSCLC recruited as part of the multicenter European Early Lung Cancer Detection Group project. A score measuring the VEGF signaling pathway was calculated by adding the tumor cell expression value of VEGF and its two receptors. The results were validated in two additional independent cohorts of patients with NSCLC.ResultsThe combination of high VEGF, VEGFR1, and VEGFR2 protein expression was associated with lower risk of disease progression in early SCC (univariate analysis, P = .008; multivariate analysis, hazard ratio, 0.62; 95% CI, 0.42 to 0.92; P = .02). The results were validated in two independent patient cohorts, confirming the favorable prognostic value of high VEGF signaling score in early lung SCC.ConclusionOur results clearly indicate that the combination of high expression of the three key elements in the VEGF pathway is associated with a good prognosis in patients with early SCC but not in patients with ADC.

Download Full-text

Novel role of p66Shc in ROS-dependent VEGF signaling and angiogenesis in endothelial cells

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.00739.2011 ◽

2012 ◽

Vol 302 (3) ◽

pp. H724-H732 ◽

Cited By ~ 53

Author(s):

Jin Oshikawa ◽

Seok-Jo Kim ◽

Eiji Furuta ◽

Cristiana Caliceti ◽

Gin-Fu Chen ◽

...

Keyword(s):

Critical Role ◽

Adaptor Protein ◽

Tube Formation ◽

Vegf Receptor ◽

Kinase Activation ◽

Vegf Signaling ◽

Rac1 Activity ◽

Interfering Rna ◽

Endothelial Growth Factor

p66Shc, a longevity adaptor protein, is demonstrated as a key regulator of reactive oxygen species (ROS) metabolism involved in aging and cardiovascular diseases. Vascular endothelial growth factor (VEGF) stimulates endothelial cell (EC) migration and proliferation primarily through the VEGF receptor-2 (VEGFR2). We have shown that ROS derived from Rac1-dependent NADPH oxidase are involved in VEGFR2 autophosphorylation and angiogenic-related responses in ECs. However, a role of p66Shc in VEGF signaling and physiological responses in ECs is unknown. Here we show that VEGF promotes p66Shc phosphorylation at Ser36 through the JNK/ERK or PKC pathway as well as Rac1 binding to a nonphosphorylated form of p66Shc in ECs. Depletion of endogenous p66Shc with short interfering RNA inhibits VEGF-induced Rac1 activity and ROS production. Fractionation of caveolin-enriched lipid raft demonstrates that p66Shc plays a critical role in VEGFR2 phosphorylation in caveolae/lipid rafts as well as downstream p38MAP kinase activation. This in turn stimulates VEGF-induced EC migration, proliferation, and capillary-like tube formation. These studies uncover a novel role of p66Shc as a positive regulator for ROS-dependent VEGFR2 signaling linked to angiogenesis in ECs and suggest p66Shc as a potential therapeutic target for various angiogenesis-dependent diseases.

Download Full-text

Impaired VEGF Signaling in Lungs with Hypoplastic Esophageal Atresia and Effects on Branching Morphogenesis

Cellular Physiology and Biochemistry ◽

10.1159/000445632 ◽

2016 ◽

Vol 39 (1) ◽

pp. 385-394 ◽

Cited By ~ 2

Author(s):

Xiaomei Liu ◽

Caixia Liu ◽

Tie Ma ◽

Yisheng Jiao ◽

Jianing Miao ◽

...

Keyword(s):

Esophageal Atresia ◽

Branching Morphogenesis ◽

Lung Growth ◽

Compensatory Response ◽

Pregnant Rats ◽

Vegf Signaling ◽

Vegf Pathway ◽

Tract Disease ◽

Endothelial Growth Factor

Background/Aims: Patients with esophageal atresia (EA) and tracheoesophageal fistula (TEF) often suffer chronic respiratory tract disease. We previously reported that primary lung maldevelopment caused by deficient branching of embryonal airways in experimental EA-TEF was induced by Adriamycin. In this study, we investigated the Vascular endothelial growth factor (VEGF) pathway in the developing lung in an EA-TEF rat model. We further analyzed the effect of recombinant VEGF treatment in vitro on branching morphogenesis of embryo lungs in experimental EA-TEF. Methods: Pregnant rats received either Adriamycin or vehicle on E7, E8 and E9. Lungs were recovered at E15, E18 and E21. Expression of VEGF and receptors (Flk-1 and Flt-1) were assessed by quantitative PCR, immunohistochemistry and immunoblotting. E13 lungs were cultured for 72 hours with 50 ng/mL of recombinant rat VEGF in serum-free medium. The rates of increase in bud count and airway contour were evaluated. Results: Our results showed a significant downregulation of VEGF during pseudoglandular and canalicular stages. In contrast, there were significantly higher levels of the Flt-1 receptor in the canalicular stage, which may represent a compensatory response to decreased VEGF. However, both variables returned to normal levels at the saccular stage. Exogenous VEGF treatment enhanced hypoplastic lung growth, evidenced by the increase in bud count and airway contour. Conclusions: A VEGF signaling defect possibly plays an important role in defective embryonic airway branching. Additionally, VEGF treatment may accelerate lung growth in EA-TEF lungs.

Download Full-text

Vascular Endothelial Growth Factor as an Immediate-Early Activator of UV-induced Skin Injury

10.1101/2020.07.13.198549 ◽

2020 ◽

Author(s):

Stella P. Hartono ◽

Victoria M. Bedell ◽

Sk. Kayum Alam ◽

Madelyn O’Gorman ◽

MaKayla Serres ◽

...

Keyword(s):

Vascular Endothelial Growth Factor ◽

Growth Factor ◽

The United States ◽

Uv Exposure ◽

Therapeutic Window ◽

Vascular Endothelial ◽

Acute Effects ◽

Vegf Signaling ◽

Vegf Pathway ◽

Endothelial Growth Factor

ABSTRACTThe negative health consequences of acute ultraviolet (UV) exposure are evident, with reports of 30,000 emergency room visits annually to treat the effects of sunburn in the United States alone. Acute effects of sunburn include erythema, edema, and severe pain, and chronic overexposure to UV radiation can lead to skin cancer. While the pain associated with the acute effects of sunburn may be relieved by current interventions, existing post-sunburn treatments are not capable of reversing the cumulative and long-term pathological effects of UV exposure, an unmet clinical need. Here we show that activation of the vascular endothelial growth factor (VEGF) pathway is a direct and immediate consequence of acute UV exposure, and activation of VEGF signaling is necessary for the initiating the acute pathological effects of sunburn. In UV-exposed human subjects, VEGF signaling is activated within hours. Topical delivery of VEGF pathway inhibitors, targeted against the ligand VEGF-A (gold nanoparticles conjugated with anti-VEGF antibodies) and small molecule antagonists of VEGF receptor signaling, prevent the development of erythema and edema in UV-exposed mice. Collectively, these findings suggest targeting VEGF signaling may reduce the subsequent inflammation and pathology associated with UV-induced skin damage, which reveals a new post-exposure therapeutic window to potentially inhibit the known detrimental effects of UV on human skin.

Download Full-text

Cooperation between VEGF and β3 integrin during cardiac vascular development

Blood ◽

10.1182/blood-2005-10-038893 ◽

2006 ◽

Vol 109 (5) ◽

pp. 1962-1970 ◽

Cited By ~ 47

Author(s):

Sara M. Weis ◽

Jeffrey N. Lindquist ◽

Leo A. Barnes ◽

Kimberly M. Lutu-Fuga ◽

Jianhua Cui ◽

...

Keyword(s):

Vascular Development ◽

Critical Role ◽

Vascular Endothelial ◽

Wild Type ◽

Vegf Signaling ◽

Cytoplasmic Vacuoles ◽

Vascular Phenotype ◽

Vessel Maturation ◽

Β3 Integrin ◽

Endothelial Growth Factor

Abstract In the developing myocardium, vascular endothelial growth factor (VEGF)–dependent neovascularization occurs by division of existing vessels, a process that persists for several weeks following birth. During this remodeling phase, mRNA expression of β3 integrin in the heart decreases significantly as vessel maturation progresses. However, in male mice lacking β3, coronary capillaries fail to mature and continue to exhibit irregular endothelial thickness, endothelial protrusions into the lumen, and expanded cytoplasmic vacuoles. Surprisingly, this phenotype was not seen in female β3-null mice. Enhanced VEGF signaling contributes to the β3-null phenotype, because these vessels can be normalized by inhibitors of VEGF or Flk-1. Moreover, intravenous injection of VEGF induces a similar angiogenic phenotype in hearts of adult wild-type mice. These findings show a clear vascular phenotype in the hearts of mice lacking β3 and suggest this integrin plays a critical role in coronary vascular development and the vascular response to VEGF.

Download Full-text

The tolerance to hypoxia is defined by a time-sensitive response of the gene regulatory network in sea urchin embryos

Development ◽

10.1242/dev.195859 ◽

2021 ◽

pp. dev.195859

Author(s):

Majed Layous ◽

Lama Khalaily ◽

Tsvia Gildor ◽

Smadar Ben-Tabou de-Leon

Keyword(s):

Early Development ◽

Sea Urchin ◽

Regulatory Networks ◽

Normal Development ◽

Axis Formation ◽

Oxygen Level ◽

Sea Urchin Embryo ◽

Tolerance To Hypoxia ◽

Vegf Pathway ◽

Gene Regulatory

Deoxygenation, the reduction of oxygen level in the oceans induced by global warming and anthropogenic disturbances, is a major threat to marine life. This change in oxygen level could be especially harmful to marine embryos that utilize endogenous hypoxia and redox gradients as morphogens during normal development. Here we show that the tolerance to hypoxic conditions changes between different developmental stages of the sea urchin embryo, possibly due to the structure of the gene regulatory networks (GRNs). We demonstrate that during normal development, bone morphogenetic protein (BMP) pathway restricts the activity of the vascular endothelial growth factor (VEGF) pathway to two lateral domains and by that controls proper skeletal patterning. Hypoxia applied during early development strongly perturbs the activity of Nodal and BMP pathways that affect VEGF pathway, dorsal-ventral (DV) and skeletogenic patterning. These pathways are largely unaffected by hypoxia applied after DV-axis formation. We propose that the use of redox and hypoxia as morphogens makes the sea urchin embryo highly sensitive to environmental hypoxia during early development, but the GRN structure provides higher tolerance to hypoxia at later stages.

Download Full-text