Non-heritable genetic diseases of the skeletal system: Pathogenesis and Treatment

骨骼系统非遗传性遗传疾病：发病机制和治疗

基本信息

批准号：
8830919
负责人：
Matthew L Warman
金额：
$ 38.83万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-04-03 至 2019-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8830919
关键词：
Ablation Adult Affect Alleles Animal Model Animals Archives Bar Codes Benign Biology Biopsy Bone Cysts Bone neoplasms Caring Cells Child Chondroblastoma Chondromyxoid Fibroma Clinical Trials Complex Computational algorithm DNA DNA Library Defect Development Diagnosis Disease Disease Progression Evaluation Face Formalin Freezing Gene Expression Profile Genes Genetic Genetic Counseling Giant Cell Tumors Glean Goals Gorham-Stout disease Health Hereditary Disease Human Individual Inherited Intervention Klippel-Trenaunay-Weber Syndrome Learning Lesion Link Lipomatosis Lymphatic Malignant - descriptor Massive Parallel Sequencing Mediating Medical Megalencephaly Morbidity - disease rate Mus Mutate Mutation Operative Surgical Procedures Other Genetics PIK3CA gene Paraffin Embedding Parents Pathogenesis Pathologic Pathologist Patients Pattern Persons Phenotype Process RNA RNA Sequences Reporter Research Research Personnel Role Sampling Sensitivity and Specificity Sequence Analysis Skeletal system Skeleton Skin Somatic Mutation Surgeon Syndrome Tamoxifen Therapeutic Tissue Sample Tissues Work alternative treatment bone loss cDNA Library cell type cohort embryonic stem cell exome genetic analysis inhibitor/antagonist insight malformation mutant novel osteosarcoma prevent recombinase reproductive research study sample collection skeletal disorder success tool transcriptome sequencing treatment strategy

项目摘要

DESCRIPTION (provided by applicant): This application focuses on the large number of children and adults who have diseases affecting the skeleton that are genetic, but not heritable. These disorders cause significant morbidity. Gorham-Stout Disease (GSD), Generalized Lymphatic Anomaly (GLA), bone cysts and tumors exemplify these diseases. The co-investigators of this application have developed an approach for identifying genetic causes for these types of disorders by combining massively parallel sequencing of DNA or RNA - recovered from a patient's affected tissues and unaffected tissues - with computational algorithms that detect somatic mosaic mutations that are either solely in sequence from affected tissue or are enriched in sequence from affected tissue compared to unaffected tissue. This approach does not rely on prospectively obtaining freshly excised lesional tissue, since it works using frozen tissue and archived formalin fixed paraffin embedded pathologic samples. We validated our approach by employing it to discover that somatic mosaic activating mutations in PIK3CA are responsible for CLOVES syndrome. We now propose to extend our work to other genetic, non-hereditary conditions. We have ascertained several well-characterized patient cohorts affected with diseases such as GSD, GLA, and bone tumors, and we have been collecting affected and unaffected tissue samples from these individuals. Our sample collection currently includes lesional tissue from 6 patients with GLA, 6 patients with GSD, 20 patients with giant cell tumors, 38 patients with chondroblastoma, 8 patients with chondromyxoid fibroma. In Aim One of this proposal, we will prepare bar-coded DNA and cDNA libraries from affected tissue and unaffected tissue, perform massively parallel sequencing, and use computational algorithms that have already enabled us to identify causative mutations in other disorders. We expect to successfully identify causative mutations in many of these new disorders. Aim Two of our proposal builds upon our discovery that PIK3CA activating mutations cause patterning defects, malformations, and overgrowths when present in somatic mosaic form. We have a mouse in which we can conditionally activate the p.1047H>R mutation, which we have found in persons with CLOVES, KTS, FIL, and isolated lymphatic malformations, and we will soon have a mouse in which we can conditionally activate and then inactivate a different disease-causing Pik3ca mutation, p.420C>R. We will use these mice to understand the pathogeneses of malformations and overgrowths that occur in PIK3CA-associated disorders and to determine whether these problems can be prevented, delayed, or reversed. Completion of these aims will link genes to phenotypes in patients whose diseases were previously impenetrable to genetic analyses and, for patients with PIK3CA-associated phenotypes, determine how lesions form and whether PIK3CA inhibition is a viable treatment strategy.

描述（由申请人提供）：此申请的重点是众多患有影响遗传骨骼但不是可遗传的骨骼的儿童和成人。这些疾病引起了明显的发病率。 Gorham-Stout疾病（GSD），广义淋巴异常（GLA），骨囊肿和肿瘤体现了这些疾病。该应用的共同评估者通过结合从患者的受影响的组织和未受影响的组织中恢复的DNA或RNA的大量平行测序来鉴定这些类型的疾病的遗传原因的方法 - 与受影响的组织中受影响的组织相称，这些算法是从患者的影响组织和未受影响的组织中恢复的方法。这种方法不依赖于前瞻性地获得新鲜切除的病变组织，因为它使用冷冻组织和存档的福尔马林固定石蜡嵌入的病理样品起作用。我们通过采用它来验证了我们的方法，发现PIK3CA中的体细胞镶嵌激活突变是丁香综合征的原因。现在，我们建议将我们的工作扩展到其他遗传，非遗传条件。我们已经确定了几个受GSD，GLA和骨肿瘤等疾病影响的良好特征的患者同类，我们一直在收集这些人的受影响和未受影响的组织样本。我们的样本收集目前包括来自6例GLA患者，6例GSD患者，20例巨细胞肿瘤患者，38例软骨细胞瘤患者，8例患有软骨瘤纤维瘤的患者的病变组织。在此提案之一中，我们将从受影响的组织和不受影响的组织中制备条形编码的DNA和cDNA文库，进行大量平行的测序，并使用已经使我们能够识别其他疾病中的病毒突变的计算算法。我们希望在许多新疾病中成功识别因果突变。目的是我们的两个提案以我们的发现为基础，即PIK3CA激活突变会导致模式缺陷，畸形和以体细节形式存在的过度生长。我们有一只小鼠，可以有条件地激活P.1047H> r突变，我们在患有丁香，KTS，FIL和孤立的淋巴畸形的人中发现，我们很快就会有条件地激活一只小鼠，然后可以灭活一个不同的疾病，可引起不同的疾病，p.420c> r。我们将使用这些小鼠了解PIK3CA相关疾病中发生的畸形和过度生长的病原体，并确定是否可以预防，延迟或逆转这些问题。这些目的的完成将把基因与以前无法接受遗传分析的疾病不可渗透的患者的表型联系起来，并且对于PIK3CA相关表型的患者，确定病变的形成以及PIK3CA抑制作用是可行的治疗策略。