Investigating the Causes of Keloid Formation

调查瘢痕疙瘩形成的原因

• 批准号: 9271039
• 负责人: Donald Alexander Glass
• 金额: $ 16.63万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-05-09 至 2021-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9271039
• 关键词: Active Learning; Affect; African; African American; Age; Applications Grants; Asians; Biometry; Candidate Disease Gene; Caucasians; Cells; Cicatrix; Clinic; Complex; Cosmetics; Coupled; DNA Sequence; Data; Data Set; Deformity; Dermatologic; Dermatology; Development; Development Plans; Disease; Enrollment; Environmental Risk Factor; Ethnic group; European; Evaluation; Event; Exons; Extended Family; Family; Family history of; Family member; Family-Based Registry; Fibroblasts; Future; Gene Expression; Generations; Genes; Genetic; Genetic Markers; Genetic Variation; Genome; Genomic DNA; Goals; Grant; High-Throughput Nucleotide Sequencing; Hispanics; Hospitals; Hypertrophic Cicatrix; Incidence; Individual; Infection; Inherited; Injury; Keloid; Link; Liquid substance; Location; Manuscripts; Medical History; Mentors; Methodology; Morbidity - disease rate; Nature; Normal tissue morphology; Northern Blotting; Operative Surgical Procedures; Participant; Pathogenesis; Phenotype; Plasma; Predisposition; Prevention; Protein Isoforms; RNA; Race; Recording of previous events; Recurrence; Registries; Research; Research Personnel; Research Training; Risk; Sampling; Series; Severities; Site; Skin; Susceptibility Gene; Testing; Time; Tissue Sample; Tissues; Training; Variant; Venipunctures; Wound Healing; Writing; admixture mapping; base; career; career development; exome; experience; genetic linkage analysis; genome database; genome wide association study; insight; logarithm; member; patient registry; population based; prevent; public health relevance; research study; response; sex; skills; transcriptome sequencing; wound; wound closure

 DESCRIPTION (provided by applicant): Keloids result from an exaggerated response to wound healing of the skin. They grow beyond the original boundaries of the injury to the skin, causing cosmetic deformities, and there is no known treatment that consistently prevents their occurrence or recurrence. The existence of families in which multiple members are affected with keloids over several generations and that certain ethnic groups are more likely to develop keloids than others imply that certain genes may predispose people to keloid formation. Surprisingly, little is known about the genetic factors contributing to keloids in individuals of African or Hispanic ancestry, which are the two ethnic groups most likely to develop keloids. There is also little known about the differences between the normal skin of those who keloid and the normal skin of those that do not. The research aims of this application will test the hypothesis that specific genetic variations confer susceptibility to keloid formation in those individuals at greatest risk: subjects of African and Hispanic ancestry. They will also test the hypothesis that there exist uncharacterized genes and gene isoforms, in keloids as well as in the normal skin of individuals that keloid that are critical to keloid pathogenesis. A keloid registry has been established to better characterize subjects and families of African and Hispanic descent with keloids. The registry has 117 families and 136 unrelated individuals with sporadic keloids to date and enrollment is ongoing. As part of enrollment a medical history, dermatologic evaluation and venipuncture are performed. Plasma and serum are aliquoted and genomic DNA is extracted and stored. In participants with a family history of keloids, the other family members (both affected and unaffected individuals) are invited to participate. Specific Aim 1 is to Identify Susceptibility Loci in Families with Keloids. Linkage analysis will be performed on all of the members of a family with familial keloids. The location of disease-causing loci within each family will be found by identifying the genetic markers that are co-inherited with the keloid phenotype. A logarithm of the odds (LOD) score will be calculated based upon the genetic markers co-segregating with the keloid phenotype within the family. A LOD score of three or more will be used to determine if a region is linked with the keloid phenotype. Candidate genes within these identified loci will be examined to identify variants that co-segregate with keloid formation. Specific Aim 2 is to Identify Genetic Variations Associated with Keloids. All exons in the genome (i.e. the exome) will be sequenced using high-throughput sequencing in selected members of families with keloids. Rare sequence variations identified in the affected family members will be further examined to identify variants that co-segregate with keloid formation. These candidate genes will be sequenced in other families that have a history of keloid formation as well as in unrelated individuals with sporadic keloids. Specific Aim 3 is t determine the differences in gene expression between keloid skin and normal skin. This aim is two-fold in nature. Sub-Aim 3.1 is to determine the differences in gene expression between keloidal fibroblasts and normal fibroblasts, while Sub-Aim 3.2 is to determine the differences in gene expression between normal fibroblasts from keloid-prone individuals and normal fibroblasts from those that do not keloid. Primary fibroblasts will be isolated from keloid tissue obtained from surgical treatments. Primary fibroblasts from perilesional normal tissue from the same individual will also be obtained. Control primary fibroblasts will be obtained from age, sex and race-matched control individuals with no personal or family history of keloids. In both sub-aims, whole transcriptome sequencing will be performed on RNA from the primary fibroblasts and differences in gene expression between the different cells will be assessed. Findings will be confirmed with Northern blot analysis and/or real-time PCR. These research aims will serve as the platform for my career development plan and will be coupled with training aims which include: 1) biostatistical analysis and methodology; 2) the analysis of high content/high complexity data sets; and 3) research management and manuscript/grant writing skills. These training aims will be achieved via didactic courses, experiential learning, and mentored studies. Together, the research and training aims of the K23 proposal will provide the training, experience, and preliminary data for future R01 grant applications. This career development plan will prepare me to become a successful independent investigator and attain my long-term career goal of becoming a national leader in keloids and genetics.

 描述(申请人提供)：瘢痕疙瘩是由于皮肤对伤口愈合的过度反应造成的。它们的生长超出了皮肤损伤的原始边界，导致了整容畸形，目前还没有已知的治疗方法来持续防止它们的发生或复发。有多名成员世代患有瘢痕疙瘩的家庭的存在，以及某些种族群体比其他民族更有可能患上瘢痕疙瘩，这意味着某些基因可能使人更容易形成瘢痕疙瘩。令人惊讶的是，人们对非洲人或西班牙人血统的人中导致瘢痕疙瘩的遗传因素知之甚少，这两个民族最有可能发生瘢痕疙瘩。瘢痕疙瘩患者的正常皮肤与非瘢痕疙瘩患者的正常皮肤之间的差异也鲜为人知。这项应用的研究目的将检验这样一个假设，即特定的基因变异使那些风险最高的人--非洲人和西班牙裔人--容易形成瘢痕疙瘩。他们还将测试这一假设，即在瘢痕疙瘩中以及在瘢痕疙瘩患者的正常皮肤中存在对瘢痕疙瘩发病至关重要的未表征的基因和基因亚型。已经建立了瘢痕疙瘩登记，以更好地描述患有瘢痕疙瘩的非洲人和西班牙裔人的对象和家庭的特征。到目前为止，登记的散发性瘢痕疙瘩有117个家庭和136个无关个人，登记工作正在进行中。作为登记病史的一部分，皮肤科评估和静脉穿刺会进行。血浆和血清被等量，基因组DNA被提取和存储。对于有瘢痕疙瘩家族史的参与者，邀请其他家庭成员(包括受影响和未受影响的个人)参加。具体目的1是确定瘢痕疙瘩家系的易感基因。对家族性瘢痕疙瘩家族的所有成员进行连锁分析。通过鉴定与瘢痕疙瘩表型共同遗传的遗传标记，可以找到每个家系中致病基因的位置。几率的对数(LOD)分数将基于与家族内的瘢痕疙瘩表型共分离的遗传标记来计算。LOD评分为3或更高将被用来确定一个区域是否与瘢痕疙瘩表型有关。将检查这些已识别基因座中的候选基因，以确定与瘢痕疙瘩形成共分离的变异。具体目标2是确定与瘢痕疙瘩相关的遗传变异。基因组中的所有外显子(即外显子)将在选定的瘢痕疙瘩家族成员中使用高通量测序进行测序。将进一步检查在受影响的家庭成员中发现的罕见序列变异，以确定与瘢痕疙瘩形成共分离的变异。这些候选基因将在其他有瘢痕疙瘩形成史的家族以及散发性瘢痕疙瘩无关的个体中进行测序。目的3：研究瘢痕疙瘩与正常皮肤基因表达的差异。这一目标具有双重性质。子目标3.1是确定瘢痕疙瘩成纤维细胞和正常成纤维细胞之间的基因表达差异，而子目标3.2是确定瘢痕疙瘩易患个体的正常成纤维细胞和非瘢痕疙瘩患者的正常成纤维细胞之间的基因表达差异。原代成纤维细胞将从手术治疗获得的瘢痕疙瘩组织中分离出来。来自同一个体的病灶周围正常组织的原代成纤维细胞也将被获得。对照原代成纤维细胞取自无瘢痕疙瘩个人或家族史的年龄、性别和种族匹配的对照个体。在这两个子目标中，将对原代成纤维细胞的RNA进行完整的转录组测序，并将评估不同细胞之间基因表达的差异。这些发现将通过Northern印迹分析和/或实时聚合酶链式反应来证实。这些研究目标将作为我职业发展计划的平台，并将与培训目标相结合，这些目标包括：1)生物统计分析和方法；2)高内容/高复杂性数据集的分析；3)研究管理和手稿/补助金写作技能。这些培训目标将通过授课课程、体验式学习和辅导性学习来实现。总之，K23提案的研究和培训目标将为未来的R01拨款申请提供培训、经验和初步数据。这份职业发展计划将为我成为一名成功的独立调查员做好准备，并实现我长期的职业目标--成为全国瘢痕疙瘩和遗传学领域的领先者。