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Engineering Growth Factor-ECM Interactions to Heal Chronic Wounds in Type 1 Diabetes

设计生长因子-ECM 相互作用来治愈 1 型糖尿病的慢性伤口

基本信息

批准号：
9037078
负责人：
JEFFREY A. HUBBELL
金额：
$ 152.65万
依托单位：
UNIVERSITY OF CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-25 至 2019-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9037078
关键词：
Affinity Amputation Amyloid beta-Protein Animals Aprotinin Autoimmune Process Becaplermin Binding Binding Sites Biocompatible Materials Biological Biological Process Biology Boxing CXCL12 gene Carboxymethylcellulose Caring Cattle Cells Chimeric Proteins Chronic Clinic Coagulation Process Complex Complication Complications of Diabetes Mellitus DTR gene Data Diabetes Mellitus Diabetic Foot Ulcer Diabetic wound Dissociation Engineering Extracellular Matrix Extracellular Matrix Proteins Factor XIIIa Fibrin Fibrin Tissue Adhesive Fibrinogen Fibronectins Gel Growth Factor Healed Heparin Binding Human Hyperglycemia Impaired wound healing Inbred NOD Mice Infiltration Insulin Insulin-Dependent Diabetes Mellitus Longevity Modeling Morbidity - disease rate Mus Non-Insulin-Dependent Diabetes Mellitus PGF gene Paper Patients Plague Protease Inhibitor Protein Engineering Proteins Proteolysis Publishing Quality of life Science Sequence Homology Serine Proteinase Inhibitors Stem cells Tenascin Therapeutic Transglutaminases Ursidae Family Variant Vascular Endothelial Growth Factors Vitronectin Work Wound Healing angiogenesis base cancer risk chemokine clinical practice cytokine diabetic wound healing experience glycemic control healing inhibitor/antagonist interest mouse model novel osteopontin platelet-derived growth factor BB public health relevance response unpublished works von Willebrand Factor wound

项目摘要

DESCRIPTION: We seek to develop and evaluate novel biomaterial/growth factor therapies to heal diabetic foot ulcers, which plague diabetes patients as a major diabetes complication, resulting in substantial morbidity including amputation and reduction of lifespan and quality of life. In current clinical practice, growth factor therapies have generally been a disappointment: Based on our published and unpublished preliminary work, we believe that the major problem with growth factor therapeutics in general is that the evolutionary context of the growth factor-extracellular matrix (ECM) microenvironment has been ignored. Our group has elucidated complex biomolecular interactions between growth factors and proteins of the ECM, which dramatically alter the biology of the growth factor. Indeed, we have come to believe that most growth factors evolved to function in concert with an ECM protein (Martino et al., Science Transl. Med. 2011, among other references). Here, we will evaluate both growth factor engineering and ECM engineering approaches directed to control the dynamic reciprocity between growth factors and the ECM in diabetic wound healing. We will on the one hand engineer growth factor variants to bind with super-affinity to the ECM, including fibrin, our selected translational biomaterial platform for ECM engineering; and we will engineer the ECM to bind with high affinity and promiscuity to growth factors. We will work in a mouse type 1 diabetes (T1D) model of delayed healing to explore responses relevant to diabetic foot ulcers, establishing a mouse clinic in which animals are treated with insulin so as to control hyperglycemia yet not restore normal euglycemia, to model imperfect glycemic control in patients. (Specific Aim 1) We will employ a protein engineering approach to engineer ECM super-affinity variants of VEGF-A, PDGF-BB, HB-EGF and CXCL12 in the NOD mouse model of delayed wound healing, targeting VEGF-A for its ability to induce angiogenesis, PDGF-BB and HB-EGF for their ability to induce re- epithelialization, and CXCL-12 for its ability to induce progenitor cell infiltration. Specifically, based on our preliminary work (Martino et al., Science 2014), a high affinity and promiscuous ECM binding domain from PlGF-2 (PlGF-2123-144) will be fused to these factors, to yield VEGF-A-PlGF-2123-144, PDGF-BB-PlGF-2123-144, HB-EGF-PlGF-2123-144 and CXCL12-PlGF-2123-144. We present preliminary data with combined VEGF-A-PlGF- 2123-144, PDGF-BB-PlGF-2123-144, dual factor therapy in a murine T2D model of diabetic wounds. (Specific Aim 2) We will do this in fibrin stabilized with a novel protease inhibitor, a human protein to replace the bovine aprotinin currently used in fibrin sealants, to avoid immunological complication upon repeated use. (Specific Aim 3) Finally, we will utilize a high affinity promiscuous growth factor binding domain that we have discovered (unpublished) in von Willebrand factor (vWF1328-1351) bound in fibrin to create high affinity binding sites to both native and engineered growth factors. Our objective is to produce a stabilized fibrin matrix, with one or perhaps two bioactive factors, to heal wounds in T1D patients.

产品说明：我们寻求开发和评估新的生物材料/生长因子疗法来治愈糖尿病足溃疡，糖尿病足溃疡作为糖尿病患者的主要并发症，导致大量发病率，包括截肢和寿命和生活质量的降低。在目前的临床实践中，生长因子治疗通常令人失望：根据我们已发表和未发表的初步工作，我们认为生长因子治疗的主要问题是生长因子-细胞外基质（ECM）微环境的进化背景被忽视了。我们的小组已经阐明了生长因子和ECM蛋白质之间复杂的生物分子相互作用，这极大地改变了生长因子的生物学特性。事实上，我们已经开始相信大多数生长因子进化为与ECM蛋白协同作用（Martino et al.，科学翻译2011，以及其他参考文献）。在这里，我们将评估两个生长因子工程和ECM工程的方法，直接控制糖尿病伤口愈合中的生长因子和ECM之间的动态相互作用。一方面，我们将工程化生长因子变体以超亲和力结合ECM，包括纤维蛋白，我们选择的ECM工程化翻译生物材料平台;我们将工程化ECM以高亲和力和混杂性结合生长因子。我们将在延迟愈合的小鼠1型糖尿病（T1 D）模型中工作，以探索与糖尿病足溃疡相关的反应，建立一个小鼠诊所，在该诊所中，动物接受胰岛素治疗，以控制高血糖，但不能恢复正常的血糖，以模拟患者的血糖控制不完善。（具体目的1）我们将采用蛋白质工程方法在延迟伤口愈合的NOD小鼠模型中工程化VEGF-A、PDGF-BB、HB-EGF和CXCL 12的ECM超亲和变体，靶向VEGF-A诱导血管生成的能力，靶向PDGF-BB和HB-EGF诱导上皮再形成的能力，靶向CXCL-12诱导血管生成的能力。祖细胞浸润。具体而言，基于我们的初步工作（Martino等人，Science 2014），来自PlGF-2（PlGF-2123-144）的高亲和力和混杂ECM结合结构域将与这些因子融合，以产生VEGF-A-PlGF-2123-144、PDGF-BB-PlGF-2123 -144、HB-EGF-PlGF-2123-144和CXCL 12-PlGF-2123-144。我们提供了在糖尿病伤口的鼠T2 D模型中使用组合的VEGF-A-PlGF-2123-144、PDGF-BB-PlGF-2123-144、双因子疗法的初步数据。（具体目标2）我们将在用新型蛋白酶抑制剂稳定的纤维蛋白中进行这一研究，该蛋白酶抑制剂是一种人类蛋白质，用于取代目前用于纤维蛋白封闭剂的牛抑肽酶，以避免重复使用时的免疫并发症。（具体目标3）最后，我们将利用我们在结合在纤维蛋白中的von Willebrand因子（vWF 1328 -1351）中发现（未公开）的高亲和力混杂生长因子结合结构域来产生对天然和工程化生长因子的高亲和力结合位点。我们的目标是生产一种稳定的纤维蛋白基质，其中含有一种或两种生物活性因子，用于T1 D患者的伤口愈合。