Heritable Disorders of Connective Tissue

结缔组织遗传性疾病

• 批准号: 10908171
• 负责人: Joan C Marini
• 金额: $ 43.76万
• 依托单位: EUNICE KENNEDY SHRIVER NATIONAL INSTITUTE OF CHILD HEALTH & HUMAN DEVELOPMENT
• 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/10908171
• 关键词: Acoustics; Adipocytes; Adult; Affect; Age; Age of Onset; Amino Acid Substitution; Antibodies; Antibody Therapy; Apoptosis; Area; Biological Assay; Body mass index; Bone Density; Bone Formation Stimulation; Bone Mineral Contents; Bone Tissue; Breathing; COL1A1 gene; COL1A2 gene; Cell Transplantation; Cells; Cephalic; Cessation of life; Chest; Chest wall structure; Child; Childhood; Clinical Research; Collaborations; Collagen; Collagen Type I; Connective Tissue; Cytoskeleton; Data; Defect; Deformity; Developmental Bone Diseases; Disease; Disease model; Dose; Dual-Energy X-Ray Absorptiometry; Family; Fatty acid glycerol esters; Femur; Fibroblasts; Fracture; Functional disorder; Gases; Gender; Genes; Genetic; Genotype; Glycine; Goals; Growth; Height; Histology; In Vitro; Individual; Intermediate Filaments; Intervention; Investigation; Knock-in; Knowledge; Laboratories; Location; Lower Extremity; Lung; Lung diseases; Mechanics; Mediating; Membrane; Metabolism; Microfilaments; Microscopy; Microtubules; Modeling; Molecular Biology; Molecular Chaperones; Molecular Genetics; Morbidity - disease rate; Mus; Mutation; National Institute of Child Health and Human Development; Natural History; Osteoblasts; Osteogenesis; Osteogenesis Imperfecta; Osteoporosis; Pain; Participant; Pathway interactions; Patients; Pattern; Peptide Hydrolases; Phenotype; Physiologic calcification; Placebo Effect; Procollagen; Property; Proteins; Pulmonary function tests; Quality of life; RNA Splicing; Randomized, Controlled Trials; Recommendation; Research; Roentgen Rays; Role; Scanning; Severities; Shapes; Site; Skin; Somatotropin; Spectroscopy, Fourier Transform Infrared; Structure of parenchyma of lung; Symptoms; Testing; Tissues; Type I Procollagen; Variant; Weight; Whole Organism; Work; bisphosphonate; bone; bone cell; bone fragility; bone mass; bone quality; clinical investigation; cortical bone; drug action; endoplasmic reticulum stress; experience; follow-up; hammerhead ribozyme; hearing impairment; heritable connective tissue disorder; improved; mineralization; mosaic; mouse model; mutant; neural; novel; pamidronate; patient variability; pediatric patients; pharmacologic; preservation; programs; pulmonary function; pulmonary function decline; response; scoliosis; side effect; skeletal; sound; spine bone structure; substantia spongiosa; translational study; transmission process; treatment group; treatment trial; young adult

In an integrated program of laboratory and clinical investigation, we study the molecular biology of the heritable connective tissue disorder osteogenesis imperfecta (OI). Our objective is to elucidate the mechanisms by which the primary gene defect causes skeletal fragility and other connective tissue symptoms and then apply the knowledge gained from our studies to the treatment of children with these conditions. Our Section has generated a knock-in murine model for OI with a COL1A1 collagen mutation, the Brtl mouse. Recently, we collaborated to investigate the material properties of Brtl cortical and endosteal bone using acoustic transmission microscopy. Collagen orientation in Brtl endosteal bone had a strong reduction in periodically alternating collagen orientation compared to WT. Also, sound velocity was significantly increased in Brtl endosteal bone, demonstrating that the predominant effect of the Brtl mutation is on endosteal bone. Brtl is also being used as the model for testing an anabolic therapy for OI, anti-sclerostin antibody (SclAB), which works by stimulating bone formation along the canonical wnt pathway. SclAB was demonstrated to be effective in increasing cortical bone formation in both young and adult Brtl mice. Brtl femora increased cortical bone formation and mechanical strength, without exacerbating the underlying brittleness of OI bone material. These data suggest SclAB treatment does not impair material properties. Although SclAB is a short-acting drug, a single dose of bisphosphonate will preserve the gains to trabecular bone mass following cessation of antibody. Recently, our collaborative studies have demonstrated that treatment of Brtl mice with SclAB does not change cranial shape in ways that would be significant for impingement of neural foramina. We identified a novel "high bone density" form of OI caused by mutations in the C-proteinase cleavage site. Children with substitutions at these residues present with fractures and a high DEXA z-score. Interestingly, despite the high DEXA, radiographs and histology are similar to type I OI and point to matrix deficiency. FTIR and BSEM revealed bone mineral content higher than other forms of OI. These data not only reveal a novel form of OI but also provide new fundamental information on roles of procollagen processing and the mechanism of tissue mineralization. We have now generated a mouse model for HBM OI, to investigate the role of type I procollagen C-propeptide cleavage in the mechanism of increased bone mineralization, both at the matrix and intracellular levels. We have also studied amino acid substitutions in the procollagen C-propeptide itself. These mutations are intriguing because they occur in a region of the promolecule that is not incorporated into matrix. On immumofluorescence microscopy, procollagen with C-propeptide defects was mislocalized to the ER lumen, in contrast to normal localization at the ER membrane. Pericellular processing of the mutant C-propeptide was defective, as were in vitro cleavage assays with purified BMP1. In bench studies aimed at understanding the basis of the phenotypic variability of patients with the identical OI-causing mutation, we collaborated on investigations of cellular cytoskeleton in Brtl lethal and surviving mice. Components of intermediate filaments, microtubules and actin filaments were all shown to be abnormal only in tissues from lethal mice. This data was extended to cells from patients with lethal and non-lethal mutations caused by identical glycine substitutions. They point to the cytoskeleton as a phenotypic modulator and potential novel target for OI treatment. We have conducted a follow-up examination to our 2007 Mutation Consortium study of genotype-phenotype relationships in classical OI. This revealed that lethality for COL1A1 mutations is associated with their location in regions important for collagen interaction with other protein, while in COL1A2, mutations resulting in splice-site variants are predominantly lethal. New modulators for phenotypic variability should be sought using murine disease models. We are continuing our clinical studies of children with types III and IV OI. The SHDBEM undertook the first randomized controlled trial of bisphosphonate in children with types III and IV OI. The treatment group experienced improvement in vertebral parameters, including BMD z-scores, central vertebral height and vertebral area, which tapered after one to two years of treatment. There was no significant change in ambulation level, lower-extremity strength or pain in treated children, which appear to be placebo effects in uncontrolled trials. We recommend that pamidronate treatment of children with types III/IV OI be limited to three years, with subsequent follow-up of bone status. Short stature is one of the cardinal features of OI. The SHDBEM has established OI-specific longitudinal growth curves for children with types III and IV OI based on data from 100 children with structural mutations in type I collagen. These curves show that the height trajectory of OI children is influenced by both type and gender, but only OI type affected the weight curves. We also derived BMI curves for OI, demonstrating distinct shifts above the CDC curves. Lung disease is the major cause of illness and death in individuals with osteogenesis imperfecta (OI), a collagen-related bone fragility disorder. Previously, external factors such as scoliosis and chest wall deformities were considered responsible for abnormal lung function in OI. Prior data from the NICHD OI Research Program showed that pulmonary function declined with age even in children who did not have scoliosis, suggesting an intrinsic component for OI lung disease. In this investigation, we studied 37 children and young adults with several types of OI, 29 of whom have structural mutations in type I collagen. Type I collagen is a component of lung parenchyma as well as the major matrix protein of bone. Study participants underwent lung function tests, X-Rays and CTs of lung tissue. Most patients had restrictive lung disease and decreased gas exchange, independent of scoliosis. Analysis of chest scans and breathing tests showed thickening of the walls of small airways and lung tissue abnormalities. We conclude that OI also causes intrinsic lung disease that negatively impacts Quality of Life (QOL) in OI. Further study of intrinsic lung abnormalities may lead to improved treatment options and improve QOL.

在实验室和临床研究的综合项目中，我们研究遗传性结缔组织疾病成骨不全症 (OI) 的分子生物学。我们的目标是阐明主要基因缺陷导致骨骼脆弱和其他结缔组织症状的机制，然后将我们研究中获得的知识应用于治疗患有这些疾病的儿童。 我们的部门已经生成了具有 COL1A1 胶原蛋白突变的成骨不全症敲入小鼠模型，即 Brtl 小鼠。最近，我们合作使用声透射显微镜研究了 Brtl 皮质骨和骨内膜骨的材料特性。与 WT 相比，Brtl 骨内膜骨中的胶原蛋白方向在周期性交替的胶原蛋白方向上显着减少。此外，Brtl 骨内骨中的声速显着增加，表明 Brtl 突变对骨内骨的主要影响。 Brtl 还被用作测试 OI 合成代谢疗法、抗硬化素抗体 (SclAB) 的模型，该抗体通过沿着规范的 WNT 途径刺激骨形成发挥作用。 SclAB 被证明可有效增加年轻和成年 Brtl 小鼠的皮质骨形成。 Brtl 股骨增加了皮质骨形成和机械强度，而不会加剧 OI 骨材料的潜在脆性。这些数据表明 SclAB 处理不会损害材料特性。尽管 SclAB 是一种短效药物，但单剂量的双膦酸盐将在停止使用抗体后保留小梁骨量的增加。最近，我们的合作研究表明，用 SclAB 治疗 Brtl 小鼠不会改变颅骨形状，从而对神经孔的冲击产生重大影响。 我们发现了一种新的“高骨密度”OI 形式，由 C 蛋白酶切割位点突变引起。这些残基发生替换的儿童会出现骨折且 DEXA z 得分较高。有趣的是，尽管 DEXA 较高，但 X 光片和组织学与 I 型 OI 相似，表明基质缺乏。 FTIR 和 BSEM 显示骨矿物质含量高于其他形式的成骨不全症。这些数据不仅揭示了成骨不全的一种新形式，而且还提供了关于前胶原加工的作用和组织矿化机制的新的基本信息。我们现在已经建立了 HBM OI 小鼠模型，以研究 I 型前胶原 C 前肽裂解在基质和细胞内水平增加骨矿化机制中的作用。 我们还研究了前胶原 C 前肽本身的氨基酸取代。这些突变很有趣，因为它们发生在未纳入基质的前分子区域。在免疫荧光显微镜下，具有 C 前肽缺陷的前胶原被错误定位到 ER 腔，这与 ER 膜上的正常定位相反。突变体 C 前肽的细胞周加工有缺陷，纯化的 BMP1 的体外切割试验也是如此。 在旨在了解具有相同 OI 突变的患者表型变异基础的实验室研究中，我们合作研究了 Brtl 致死和存活小鼠的细胞骨架。仅在致死小鼠的组织中，中间丝、微管和肌动蛋白丝的成分均显示异常。该数据扩展到来自具有相同甘氨酸取代引起的致死和非致死突变的患者的细胞。他们指出细胞骨架是一种表型调节剂和成骨不全治疗的潜在新靶点。 我们对 2007 年突变联盟关于经典 OI 基因型-表型关系的研究进行了后续检查。这表明，COL1A1 突变的致命性与其所在区域对胶原蛋白与其他蛋白质相互作用很重要，而在 COL1A2 中，导致剪接位点变异的突变主要是致命的。应使用小鼠疾病模型寻找新的表型变异调节剂。 我们正在继续对 III 型和 IV 型 OI 儿童进行临床研究。 SHDBEM 开展了第一个双磷酸盐治疗 III 型和 IV 型 OI 儿童的随机对照试验。治疗组的椎骨参数有所改善，包括 BMD z 评分、椎体中心高度和椎体面积，这些参数在治疗一到两年后逐渐缩小。接受治疗的儿童的行走水平、下肢力量或疼痛没有显着变化，这在非对照试验中似乎是安慰剂效应。我们建议 III/IV 型 OI 儿童的帕米膦酸钠治疗期限为三年，并随后随访骨状况。 身材矮小是成骨不全症的主要特征之一。 SHDBEM 根据 100 名 I 型胶原蛋白结构突变儿童的数据，建立了 III 型和 IV 型 OI 儿童的 OI 特异性纵向生长曲线。这些曲线表明，成骨不全儿童的身高轨迹同时受到类型和性别的影响，但只有成骨不全类型影响体重曲线。我们还得出了 OI 的 BMI 曲线，显示出高于 CDC 曲线的明显变化。 肺部疾病是成骨不全症（OI）患者患病和死亡的主要原因，OI 是一种与胶原蛋白相关的骨脆性疾病。此前，脊柱侧凸和胸壁畸形等外部因素被认为是成骨不全肺功能异常的原因。 NICHD OI 研究计划的先前数据显示，即使对于没有脊柱侧凸的儿童，肺功能也会随着年龄的增长而下降，这表明 OI 肺部疾病的内在因素。在这项调查中，我们研究了 37 名患有多种类型成骨不全症的儿童和年轻人，其中 29 名患者的 I 型胶原蛋白存在结构突变。 I型胶原蛋白是肺实质的组成部分，也是骨的主要基质蛋白。研究参与者接受了肺功能测试、肺组织 X 光检查和 CT 检查。大多数患者患有限制性肺部疾病和气体交换减少，与脊柱侧凸无关。胸部扫描和呼吸测试分析显示小气道壁增厚和肺组织异常。 我们得出的结论是，成骨不全还会导致内在的肺部疾病，从而对成骨不全患者的生活质量 (QOL) 产生负面影响。对内在肺部异常的进一步研究可能会改善治疗方案并提高生活质量。

项目成果

Pulmonary function and structure abnormalities in children and young adults with osteogenesis imperfecta point to intrinsic and extrinsic lung abnormalities.

患有成骨不全症的儿童和年轻人的肺功能和结构异常指向内在和外在的肺异常。

• DOI: 10.1136/jmg-2022-109009
• 发表时间: 2023
• 期刊: Journal of medical genetics
• 影响因子: 4
• 作者: Gochuico,BernadetteR;Hossain,Mahin;Talvacchio,SaraK;Zuo,MeiXingG;Barton,Mark;DangDo,AnNgoc;Marini,JoanC
• 通讯作者: Marini,JoanC

Longitudinal growth curves for children with classical osteogenesis imperfecta (types III and IV) caused by structural pathogenic variants in type I collagen.

• DOI: 10.1038/s41436-018-0307-y
• 发表时间: 2019-05
• 期刊: Genetics in medicine : official journal of the American College of Medical Genetics
• 作者: Barber LA;Abbott C;Nakhate V;Do AND;Blissett AR;Marini JC
• 通讯作者: Marini JC

4-PBA ameliorates cellular homeostasis in fibroblasts from osteogenesis imperfecta patients by enhancing autophagy and stimulating protein secretion.

• DOI: 10.1016/j.bbadis.2018.02.002
• 发表时间: 2018-05
• 期刊: Biochimica et biophysica acta. Molecular basis of disease
• 作者: Besio R;Iula G;Garibaldi N;Cipolla L;Sabbioneda S;Biggiogera M;Marini JC;Rossi A;Forlino A
• 通讯作者: Forlino A