Vacular type of Ehlers-Danlos Syndrome: experimental models and treatment

血管型埃勒斯-当洛斯综合征：实验模型和治疗

• 批准号: 7732334
• 负责人: Mark Talan
• 金额: $ 55.02万
• 依托单位: NATIONAL INSTITUTE ON AGING
• 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/7732334
• 关键词: Affect; Age; Alleles; Animals; Aorta; Attenuated; Biomechanics; Blood Vessels; Caliber; Cell Proliferation; Collagen; Colon; Condition; Development; Disease; Ehlers-Danlos Syndrome; Elastin; Evaluation; Experimental Models; Family history of; Female; Fibrosis; Frequencies; Genes; Genetic Models; Genotype; Genus Cola; Glycine; Goals; High Pressure Liquid Chromatography; Histology; Inborn Genetic Diseases; Individual; Inflammation; Intervention; Intestines; Joint Laxity; Lead; Legal patent; Life Expectancy; Measures; Modality; Modeling; Molecular Genetics; Mus; Mutation; Nature; Nucleotides; Numbers; Operative Surgical Procedures; Peptides; Procollagen Type III; Property; Proteins; Reverse Transcriptase Polymerase Chain Reaction; Risk; Rupture; Skin; Testing; Therapeutic; Time; Tissues; Ultrasonography; Uterine Rupture; Wild Type Mouse; disease classification; falls; heritable connective tissue disorder; in vivo; male; mortality; pre-clinical; prenatal; pressure; prevent; programs; prophylactic; research clinical testing; research study; tissue culture; tool

The broad objective of this program is to perform preclinical experimentation on the mouse genetic models of VEDS to test the efficacy of different therapeutic modalities to attenuate the vascular fragility and, thus, to prevent or to reduce the risk of vascular complications. With repect to specific steps outlined in Objectives, this year we concentrated on steps I and III. Characterization of existing genetic model of VEDS The haploinsufficiency for one COL3A1 allele is one of the genotypes resulting in VEDS. Homozygous Col3a1 mice, the only currently available and described model of VEDS, cannot be used for experiments due to extremely high prenatal mortality. We hypothesized, that heterozygous Col3a1 mice, originally described as phenotypically normal, can serve as an experimental model of haploinsufficiency. We compared aortas and colons of 9, 14 and 21 months old (+/-) and (+/+) Col3a1 mice in vivo (high frequency sonography and pressure-volume analyses) and ex vivo (histology and biomechanical properties). In all ages the aorta in +/- mice had a lower compliance, larger lumen diameter, and lower ex vivo rupture pressure, than in wild type mice. Histological evaluation of aortas of +/- mice demonstrated abnormalities among 100% of male and 50% of female mice, revealing elastin fragmentation, spindle cell proliferation, inflammation, and reactive fibrosis. The colon of +/- animals had higher compliance and lower maximal (pre-rupture) pressure (higher fragility) in 9-21 months old animals. This was associated with a lower collagen III content detected by quantitative RT-PCR and, on protein level, with peptides separated by HPLC and detected with LC-MS. Thus, the +/- Col3A1 mouse could serve as an experimental model for the VEDS. Development of molecular genetic tools to suppress specific mutations in tissue culture experiments The experiments conducted during this year led to a patent application, which is currently being filed, and thus cannot be disclosed at this time.

该计划的主要目的是对VEDS的小鼠遗传模型进行临床前实验，以测试不同治疗方式减轻血管脆性的有效性，从而预防或降低血管并发症的风险。关于目标中概述的具体步骤，今年我们集中注意步骤一和步骤三。 VEDS现有遗传模型的表征 一个COL 3A 1等位基因的单倍不足是导致VEDS的基因型之一。 纯合子Col 3a 1小鼠，目前唯一可用和描述的VEDS模型，不能用于实验，由于极高的产前死亡率。我们假设，最初被描述为表型正常的杂合子Col 3a 1小鼠可以作为单倍不足的实验模型。 我们比较了9、14和21月龄（+/-）和（+/+）Col 3a 1小鼠体内（高频超声和压力-容积分析）和体外（组织学和生物力学特性）的结肠和结肠。 在所有年龄段中，+/-小鼠的主动脉具有比野生型小鼠更低的顺应性、更大的管腔直径和更低的离体破裂压。+/-小鼠的结缔组织的组织学评价表明，100%的雄性和50%的雌性小鼠中存在异常，揭示了弹性蛋白断裂、梭形细胞增殖、炎症和反应性纤维化。在9-21月龄动物中，+/-动物的结肠具有较高的顺应性和较低的最大（破裂前）压力（较高的脆性）。这与通过定量RT-PCR检测到的较低的III型胶原含量相关，并且在蛋白质水平上，与通过HPLC分离并通过LC-MS检测到的肽相关。因此，+/-Col 3A 1小鼠可以作为VEDS的实验模型。 开发分子遗传工具抑制组织培养实验中的特定突变 今年进行的实验导致了一项专利申请，目前正在提交，因此目前无法披露。