Role Of Extracellular Matrix In Cell Differentiation And

细胞外基质在细胞分化中的作用

• 批准号: 7317791
• 负责人: HYNDA K KLEINMAN
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF DENTAL & CRANIOFACIAL RESEARCH
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7317791
• 关键词: Role; Extracellular; Matrix; Cell; Differentiation

The extracellular matrix is important in embryogenesis and in tissue repair. From in vitro studies using purified components, a better understanding of how cells adhere, migrate, proliferate, and differentiate in response to tissue- and cell-specific matrix molecules has been established. We have found that the basement membrane, the extracellular matrix which underlies all epithelial cells and endothelial cells and surrounds nerve cells, promotes cell differentiation in vitro. When cultured on basement membrane, endothelial cells form capillary-like structures with a lumen, chondrocytes form cartilage, salivary cells form glands, etc. Our goal is to define the molecular and cellular events involved in this process. Our approach has been (1) to identify the biologically active matrix components, with respect to cell adhesion, migration, proliferation, and differentiation, (2) localize active sites on the matrix component with synthetic peptides, (3) identify and characterize cellular receptors, (4) gain an understanding of the intracellular events involved in the biological response, and (5) identify genes induced by the extracellular matrix. We work in several in vitro model systems including primary and established breast and melanoma tumor cells, endothelial cells, and salivary glands and cells. We have used an endothelial cell tube assay in vitro as well as in vivo angiogenesis assays to define molecules that regulate vessel formation in development, repair, and disease. Our goal is to discover new angiogenic regulators that have physiological and clinical relevance. We have worked with the basement membrane protein laminin and identified active sites and cellular receptors for cell adhesion and angiogenesis. Some of these active peptides are being developed into slow-release scaffolds for further in vivo testing for wound healing in the skin and in the oral cavity. We have also focused on the thymosin beta-4 gene, which is induced by endothelial cells as they differentiate into capillary-like structures on a basement membrane substratum. We find that thymosin beta-4 promotes dermal wound repair via increased angiogenesis and keratinocyte cell migration. It promotes wound repair in normal animals as well as in animals with delayed wound healing, including diabetic and aged animals. Thymosin beta-4 also promotes hair growth in aged animals and in animals treated with cyclophosphamide, which is a model for chemotherapy-induced hair loss. Hair growth is also stimulated in athymic nude mice, which lack abundant hair shafts. We have identified the active site on this protein to a 7-amino acid actin binding domain. The cellular receptor may be surface actin. We now have created a transgenic mouse that overexpresses this protein in its skin. The mouse appears to have accelerated wound healing and hair regrowth after shaving. Unexpectedly, the transgenic mouse also has abnormal tooth development, which we are currently investigating. This protein is currently in phase 2 human clinical trials for wound healing for diabetic and aged patients and for epidermolysis bullosa patients. Clinical trials for eye repair will begin soon. Such trials are conducted by a company that has licensed the NIH patent for this use. Our goal is to understand how the extracellular matrix regulates tissue formation, repair, and certain pathological processes.

细胞外基质在胚胎发生和组织修复中是重要的。从使用纯化组分的体外研究中，已经建立了对细胞如何响应组织和细胞特异性基质分子而粘附、迁移、增殖和分化的更好理解。我们已经发现，基底膜，细胞外基质的基础上所有的上皮细胞和内皮细胞和周围的神经细胞，促进细胞分化在体外。当培养在基底膜上时，内皮细胞形成具有管腔的毛细血管样结构，软骨细胞形成软骨，唾液细胞形成腺体等。我们的目标是确定参与这一过程的分子和细胞事件。我们的方法是（1）鉴定与细胞粘附、迁移、增殖和分化有关的生物活性基质组分，（2）用合成肽定位基质组分上的活性位点，（3）鉴定和表征细胞受体，（4）了解生物反应中涉及的细胞内事件，和（5）鉴定细胞外基质诱导的基因。我们在几个体外模型系统中工作，包括原发性和已建立的乳腺和黑色素瘤肿瘤细胞，内皮细胞，唾液腺和细胞。我们已经使用了体外内皮细胞管测定以及体内血管生成测定来定义在发育、修复和疾病中调节血管形成的分子。我们的目标是发现具有生理和临床相关性的新的血管生成调节因子。我们研究了基底膜蛋白层粘连蛋白，并确定了细胞粘附和血管生成的活性位点和细胞受体。这些活性肽中的一些正在被开发成缓释支架，用于皮肤和口腔中伤口愈合的进一步体内测试。 我们也集中在胸腺素β-4基因，这是诱导内皮细胞，因为他们分化成毛细血管样结构的基底膜基质。我们发现胸腺素β-4通过增加血管生成和角质形成细胞迁移促进皮肤伤口修复。它促进正常动物以及伤口愈合延迟的动物（包括糖尿病和老年动物）的伤口修复。胸腺素β-4还促进老年动物和环磷酰胺治疗动物的毛发生长，环磷酰胺是化疗诱导的脱发模型。在无胸腺的裸鼠中，毛发生长也受到刺激，因为它们缺乏丰富的毛干。我们已经确定该蛋白的活性位点为7个氨基酸的肌动蛋白结合结构域。细胞受体可以是表面肌动蛋白。我们现在已经创造了一种在皮肤中过度表达这种蛋白质的转基因小鼠。小鼠似乎在剃毛后加速了伤口愈合和毛发再生。出乎意料的是，转基因小鼠也有异常的牙齿发育，我们目前正在调查。这种蛋白质目前正在进行2期人体临床试验，用于糖尿病和老年患者以及大疱性表皮病患者的伤口愈合。眼部修复的临床试验将很快开始。这些试验是由一家公司进行的，该公司已授权NIH专利用于此用途。我们的目标是了解细胞外基质如何调节组织形成，修复和某些病理过程。

项目成果

Cyclic peptides from the loop region of the laminin alpha 4 chain LG4 module show enhanced biological activity over linear peptides.

• DOI: 10.1021/bi049009u
• 发表时间: 2004-10
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: F. Yokoyama;N. Suzuki;M. Haruki;N. Nishi;S. Oishi;N. Fujii;A. Utani;H. Kleinman;M. Nomizu

Selective side-chain modification of cysteine and arginine residues blocks pathogenic activity of HIV-1-Tat functional peptides.

半胱氨酸和精氨酸残基的选择性侧链修饰可阻断 HIV-1-Tat 功能肽的致病活性。

• DOI: 10.1016/j.peptides.2005.09.013
• 发表时间: 2006
• 期刊: Peptides
• 影响因子: 3
• 作者: Devadas,Krishnakumar;Boykins,RobertA;Hardegen,NeilJ;Philp,Deborah;Kleinman,HyndaK;Osa,Etin-Osa;Wang,Jiun;Clouse,KathleenA;Wahl,LarryM;Hewlett,IndiraK;Rappaport,Jay;Yamada,KennethM;Dhawan,Subhash
• 通讯作者: Dhawan,Subhash

Metallothionein promotes laminin-1-induced acinar differentiation in vitro and reduces tumor growth in vivo.

• 发表时间: 2002-09
• 期刊: Cancer research
• 影响因子: 11.2
• 作者: D. Hecht;D. Jung;V. Prabhu;P. Munson;M. Hoffman;H. Kleinman