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，具有快速血管形成的独特生命周期 在婴儿期的前 6-9 个月内，随后经过数年的时间血管缓慢自发复旧 年。对于大多数儿童来说，IH 不会构成严重威胁，无需治疗；然而，大约在 10% 的病例中，IH 会急剧增大，威胁器官并导致永久性毁容。过去的 10年来，普萘洛尔，一种著名的非选择性β-肾上腺素受体拮抗剂，已成为一线药物 危及 IH 的治疗方法，但它如何以及为何在减少 IH 血管过度生长方面如此有效？ 仍然是一个谜。由于高达 18% 的 IH 失败，因此迫切需要改进普萘洛尔治疗 为了做出反应，停药后高达 25% 的婴儿会恢复生长，并且 37% 的接受普萘洛尔治疗的婴儿需要 在 5-6 岁时进行手术，以尽量减少残留纤维脂肪残留引起的畸形。为了改进 普萘洛尔，有必要阐明其抗血管过度生长的作用机制，然后 为推进 IH 药物治疗以及其他潜在的新生血管疾病提供了一条前进之路。 在之前的资助周期中，我们从人类 IH 手术标本中鉴定出了血管瘤干细胞 (HemSC) 可以分化为内皮细胞、周细胞和脂肪细胞，并在内部形成血管瘤样血管 植入免疫缺陷小鼠体内需要 7 天。我们实验室和其他实验室的后续研究验证了 HemSC 作为 IH 启动细胞。我们最近的结果表明，转录的小分子抑制剂 SOX18 因子和普萘洛尔均能有效阻止 HemSC 向内皮细胞分化。此外， 普萘洛尔的 R(+) 对映体缺乏 β-肾上腺素能受体拮抗活性，同样有效。 这一新发现确定了一种不依赖 β-肾上腺素受体、SOX18 依赖的机制，通过该机制 普萘洛尔可减少 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 将使用我们的体内模型直接、严格地测试 IH 血管形成中对 SOX18 的要求 IH 衍生的 HemSC 在裸鼠体内形成类似 IH 的血管。目标 2 将调查二聚化状态 IH 中的 SOX18（子目标 2a），普萘洛尔和 R(+) 对映体如何破坏 SOX18 二聚化和子目标 细胞定位（子目标 2b），以及它如何改变转录以防止 HemSC 血管形成 （分目标 2c）。并行进行的目标 3 将使用以下方法分析我们现有的下一代测序数据： 新的生物信息学工具可识别潜在的染色体易位或小拷贝数变异 可以产生具有新活性的融合转录本（子目标 3a），并将在 IH 组织和新鲜分离的 IH 细胞作为鉴定融合转录本的替代方法（子目标 3b）；一次 确定后，将在体外和体内模型中测试与 SOX18 和 IH 血管形成的联系。

项目成果

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

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

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

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

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

血管瘤干细胞的粘附和血管生成潜力需要α6-整合素。

• 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