Blood vessel assembly from multipotent hemangioma-derived stem cells

来自多能血管瘤干细胞的血管组装

• 批准号: 10609870
• 负责人: Joyce E. Bischoff
• 金额: $ 49.42万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10609870
• 关键词: 6 year old; Adipocytes; Adrenergic Antagonists; Benign; Binding; Blood Vessels; Breathing; Cell Separation; Cells; Child; Chromatin; Chromosomal translocation; Copy Number Polymorphism; Corneal Neovascularization; Data; Deformity; Didelphidae; Dimerization; Dominant-Negative Mutation; Endothelial Cells; Endothelium; Face; Funding; Gene Fusion; Genes; Genetic; Genetic Transcription; Goals; Growth; Hemangioendothelioma; Hemangioma; Home; Homo; Human; Hypotrichosis; Immune; Implant; In Vitro; Infant; Kidney; Learning; Life Cycle Stages; Location; Lymphedema; Medical; Methods; Modeling; Molecular; Mus; Mutation; Neoplasms in Vascular Tissue; Nuclear Extract; Nude Mice; Operative Surgical Procedures; Organ; Pathogenesis; Pathologic; Pathway interactions; Patients; Pericytes; Pharmaceutical Preparations; Prevalence; Progress Reports; Propranolol; Publications; Reagent; Role; SOX18 gene; Sampling; Somatic Mutation; Specimen; Strawberry nevus; Structure; Syndrome; Telangiectasis; Testing; Time; Tissues; Transcript; Variant; Vascular Diseases; Vascular Proliferation; Visual impairment; Work; Xenograft Model; beta-adrenergic receptor; bioinformatics tool; blood vessel development; dimer; effective therapy; enantiomer; endothelial stem cell; exome; experimental study; feeding; genome sequencing; human stem cells; improved; in vivo Model; infancy; insight; knock-down; neovascular; neovascularization; next generation sequencing; novel; pharmacologic; prevent; small hairpin RNA; small molecule inhibitor; stem cells; transcription factor; transcriptome sequencing; whole genome

Project Abstract Infantile hemangioma (IH) is a common vascular tumor with a unique lifecycle of rapid blood vessel formation over the first 6-9 months of infancy, followed by a slow spontaneous involution of blood vessels over several years. For most children, IH does not pose a serious threat and therapy is unnecessary; however, in about 10% of cases, IH can enlarge dramatically, threaten organs and cause permanent disfigurement. Over the last 10 years, propranolol, a well-known non-selective β-adrenergic receptor antagonist, has emerged as first-line therapy for endangering IH, yet how and why it works so well in reducing the vascular overgrowth in IH has remained a mystery. There is a significant need to improve propranolol therapy because up to 18% of IHs fail to respond, up to 25% resume growth when the drug is stopped, and 37% of propranolol-treated infants require surgery at 5-6 years of age to minimize deformity caused by remaining fibrofatty residua. To improve on propranolol, it is essential to elucidate it’s mechanism of action against vascular overgrowth, which will then provide a path forward to advance IH medical therapy, and potentially other neovascular diseases as well. In previous funding cycles, we identified a hemangioma stem cell (HemSC) from human IH surgical specimens that can differentiate into endothelial cells, pericytes and adipocytes and form hemangioma-like vessels within 7 days when implanted into immune-deficient mice. Subsequent studies from our lab and others validate HemSCs as the IH-initiating cell. Our recent results show that a small molecule inhibitor of the transcription factor SOX18 and propranolol both effectively block HemSC-to-endothelial differentiation. Furthermore, the R(+) enantiomer of propranolol, which lacks β-adrenergic receptor antagonistic activity, is equally effective. This novel discovery identifies a β-adrenergic receptor-independent, SOX18-dependent mechanism by which propranolol reduces vascular overgrowth in IH. To investigate deeply, we propose three specific aims. Aim 1 will directly and rigorously test the requirement for SOX18 in IH vessel formation using our in vivo model in which IH-derived HemSC form IH-like blood vessels in nude mice. Aim 2 will investigate dimerization status of SOX18 in IH (sub-aim 2a), how propranolol and the R(+) enantiomer disrupt SOX18 dimerization and sub- cellular localization (sub-aim 2b), and how this alters transcription to prevent HemSC-blood vessel formation (sub-aim 2c). Aim 3, conducted in parallel, will analyze our existing next generation sequencing data using new bioinformatic tools to identify potential chromosomal translocations or small copy number variants that could produce fusion transcripts with new activities (sub-aim 3a) and will perform deep coverage RNA-Seq on IH tissue and freshly isolated IH cells as an alternative method to identify fusion transcripts (sub-aim 3b); once identified, the connection to SOX18 and IH vessel formation will tested in in vitro and in vivo models.

项目摘要 婴儿血管瘤（IH）是一种常见的血管肿瘤，具有独特的快速血管形成生命周期 在最初的6-9个月中 年。对于大多数儿童而言，IH不会构成严重的威胁，并且没有必要。但是，大约 IH有10％的病例可以大大扩大，威胁器官并导致永久毁容。最后 10年，普遍是一种众所周知的非选择性β-肾上腺素受体拮抗剂，已成为第一线 危害IH的疗法，但是如何以及为什么在减少IH的血管过度生长方面如此奏效 仍然是一个神秘的。由于多达18％的IH失败 为了做出反应，停止药物时最多恢复增长，而37％的普遍治疗的婴儿需要 5-6岁的手术可最大程度地减少剩余的纤维f脂残留物引起的畸形。进步 普萘洛尔，必须阐明其针对血管过度生长的作用机理，然后将 为进步IH医疗疗法以及其他可能的其他新生血管疾病提供了前进的途径。 在以前的资金周期中，我们从人IH手术标本中鉴定出一个血管瘤干细胞（HEMSC） 可以区分内皮细胞，周细胞和脂肪细胞，并形成类似血管瘤的血管 7天植入免疫缺陷小鼠。我们实验室和其他人的随后研究验证 HEMSC作为IH发射细胞。我们最近的结果表明，转录的小分子抑制剂 因子Sox18和普萘洛尔都有效地阻断了HEMSC至内皮分化。此外， 缺乏β-肾上腺素受体拮抗活性的普萘洛尔的R（+）对映异构体同样有效。 这个新颖的发现鉴定了β-肾上腺素接收器独立的SOX18依赖性机制 普萘洛尔减少了IH中的血管过度生长。为了深入研究，我们提出了三个具体目标。目标1 将直接，严格地测试使用我们的体内模型在IH容器形成中Sox18的要求 IH衍生的HEMSC在裸鼠中形成了IH样血管。 AIM 2将调查二聚化状态 IH（sub-aim 2a）中的sox18，普萘洛尔和r（+）参与者如何破坏sox18二聚体和子 - 细胞定位（Sub-aim 2b），以及该如何改变转录以防止Hemsc血管形成 （sub-aim 2c）。 AIM 3并行进行，将使用 新的生物信息学工具，以识别潜在的染色体易位或小型拷贝数变体 可以通过新活动（sub-aim 3a）产生融合成绩单，并将在 IH组织和新鲜分离的IH细胞是鉴定融合转录本的替代方法（Sub-iaM 3b）；一次 鉴定出，与Sox18和IH血管形成的连接将在体外和体内模型中进行测试。

项目成果

IGF-2 and FLT-1/VEGF-R1 mRNA levels reveal distinctions and similarities between congenital and common infantile hemangioma.

IGF-2 和 FLT-1/VEGF-R1 mRNA 水平揭示了先天性血管瘤和普通婴儿血管瘤之间的区别和相似之处。

• DOI: 10.1203/pdr.0b013e318163a243
• 发表时间: 2008
• 期刊: Pediatric research
• 影响因子: 3.6
• 作者: Picard,Arnaud;Boscolo,Elisa;Khan,ZiaA;Bartch,TatiannaC;Mulliken,JohnB;Vazquez,MariePaule;Bischoff,Joyce
• 通讯作者: Bischoff,Joyce

Endoglin regulates mural cell adhesion in the circulatory system.

• DOI: 10.1007/s00018-015-2099-4
• 发表时间: 2016-04
• 期刊: Cellular and molecular life sciences : CMLS
• 作者: Rossi E;Smadja DM;Boscolo E;Langa C;Arevalo MA;Pericacho M;Gamella-Pozuelo L;Kauskot A;Botella LM;Gaussem P;Bischoff J;Lopez-Novoa JM;Bernabeu C
• 通讯作者: Bernabeu C

α6-Integrin is required for the adhesion and vasculogenic potential of hemangioma stem cells.

• DOI: 10.1002/stem.1539
• 发表时间: 2014-03
• 期刊: STEM CELLS
• 影响因子: 5.2
• 作者: Smadja, David M.;Guerin, Coralie L.;Boscolo, Elisa;Bieche, Ivan;Mulliken, John B.;Bischoff, Joyce
• 通讯作者: Bischoff, Joyce

Rapamycin suppresses self-renewal and vasculogenic potential of stem cells isolated from infantile hemangioma.

• DOI: 10.1038/jid.2011.300
• 发表时间: 2011-12
• 期刊: JOURNAL OF INVESTIGATIVE DERMATOLOGY
• 影响因子: 6.5
• 作者: Greenberger, Shoshana;Yuan, Siming;Walsh, Logan A.;Boscolo, Elisa;Kang, Kyu-Tae;Matthews, Benjamin;Mulliken, John B.;Bischoff, Joyce
• 通讯作者: Bischoff, Joyce

Glucose transporter 1-positive endothelial cells in infantile hemangioma exhibit features of facultative stem cells.

• DOI: 10.1002/stem.1841
• 发表时间: 2015-01
• 期刊: STEM CELLS
• 影响因子: 5.2
• 作者: Huang, Lan;Nakayama, Hironao;Klagsbrun, Michael;Mulliken, John B.;Bischoff, Joyce
• 通讯作者: Bischoff, Joyce