Investigating the Causes of Keloid Formation

调查瘢痕疙瘩形成的原因

• 批准号: 9913469
• 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 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9913469
• 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; 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; Skin wound healing; Susceptibility Gene; Testing; Time; Tissue Sample; Tissues; Training; Variant; Venipunctures; Writing; admixture mapping; base; career; career development; dbSNP; 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; wound healing

 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（由申请人提供）：疤痕疙瘩是由皮肤伤口愈合过度反应引起的。它们生长超出了皮肤损伤的原始边界，导致外观畸形，并且没有已知的治疗方法可以始终防止它们的发生或复发。多个家庭成员在几代人中都受到疤痕疙瘩的影响，并且某些种族比其他人更容易患疤痕疙瘩，这意味着某些基因可能使人们容易形成疤痕疙瘩。令人惊讶的是，对于非洲或西班牙血统的个体中导致疤痕疙瘩的遗传因素知之甚少，这两个种族最有可能患疤痕疙瘩。对于患有疤痕疙瘩的人和没有疤痕疙瘩的人的正常皮肤之间的差异也知之甚少。本申请的研究目的将检验以下假设：特定的遗传变异使那些风险最大的个体（非洲和西班牙血统的受试者）对疤痕疙瘩形成易感性。他们还将测试这样的假设：在疤痕疙瘩以及疤痕疙瘩个体的正常皮肤中存在对疤痕疙瘩发病机制至关重要的未表征的基因和基因亚型。 已经建立了疤痕疙瘩登记处，以更好地描述患有疤痕疙瘩的非洲和西班牙裔受试者和家庭的特征。迄今为止，该登记处已有 117 个家庭和 136 名无亲属关系的个体患有散发性瘢痕疙瘩，登记工作正在进行中。作为登记的一部分，需要进行病史、皮肤病学评估和静脉穿刺。将血浆和血清等分，提取并储存基因组 DNA。对于有疤痕疙瘩家族史的参与者，其他家庭成员（受影响的和未受影响的个人）被邀请参加。 具体目标 1 是确定瘢痕疙瘩家族的易感位点。将对患有家族性瘢痕疙瘩的所有家庭成员进行连锁分析。通过鉴定与疤痕疙瘩表型共同遗传的遗传标记，可以找到每个家族中致病位点的位置。将根据与家族内瘢痕疙瘩表型共分离的遗传标记来计算对数几率 (LOD) 分数。 3 或更高的 LOD 评分将用于确定某个区域是否与疤痕疙瘩表型相关。将检查这些已识别基因座内的候选基因，以确定与疤痕疙瘩形成共分离的变异。 具体目标 2 是识别与疤痕疙瘩相关的遗传变异。基因组中的所有外显子（即外显子组）将使用高通量测序对选定的瘢痕疙瘩家族成员进行测序。在受影响的家族成员中发现的罕见序列变异将被进一步检查，以识别与疤痕疙瘩形成共分离的变异。这些候选基因将在具有疤痕疙瘩形成史的其他家族以及患有散发性疤痕疙瘩的无关个体中进行测序。 具体目标3是确定瘢痕疙瘩皮肤和正常皮肤之间基因表达的差异。这一目标本质上有两个目的。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基因表达的差异，而子目标3.2是确定来自有瘢痕疙瘩倾向个体的正常成纤维细胞和来自不患有瘢痕疙瘩的正常成纤维细胞之间基因表达的差异。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。还将获得来自同一个体的病灶周围正常组织的原代成纤维细胞。对照原代成纤维细胞将从年龄、性别和种族匹配的对照个体中获得，且没有疤痕疙瘩个人史或家族史。在这两个子目标中，将对原代成纤维细胞的 RNA 进行全转录组测序，并评估不同细胞之间基因表达的差异。结果将通过 Northern 印迹分析和/或实时 PCR 得到证实。 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，其中包括：1）生物统计分析和方法； 2）高内容/高复杂度数据集的分析； 3) 研究管理和稿件/资助金写作技巧。这些培训目标将通过教学课程、体验式学习和指导研究来实现。 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。这个职业发展计划将使我做好准备，成为一名成功的独立研究者，并实现成为疤痕疙瘩和遗传学领域的全国领导者的长期职业目标。

项目成果

Plasma Angiotensin-converting Enzyme Levels in Patients With Keloids and/or Hypertension.

瘢痕疙瘩和/或高血压患者的血浆血管紧张素转换酶水平。

• 发表时间: 2018
• 期刊: Wounds : a compendium of clinical research and practice
• 作者: Stewart,Jacob;Glass2nd,DonaldA
• 通讯作者: Glass2nd,DonaldA

Pentoxifylline for the Prevention of Postsurgical Keloid Recurrence.

己酮可可碱用于预防术后瘢痕疙瘩复发。

• DOI: 10.1097/dss.0000000000002090
• 发表时间: 2020
• 期刊: Dermatologic surgery : official publication for American Society for Dermatologic Surgery [et al.]
• 作者: Tan,Andrea;MartinezLuna,Orlando;Glass2nd,DonaldA
• 通讯作者: Glass2nd,DonaldA

Atopic dermatitis is associated with an increased risk of keloids: A case-control study.

特应性皮炎与疤痕疙瘩风险增加相关：一项病例对照研究。

• DOI: 10.1016/j.jaad.2022.12.013
• 发表时间: 2023
• 期刊: Journal of the American Academy of Dermatology
• 影响因子: 13.8
• 作者: Limmer,EmilyE;Knowles,Ariel;Deng,Junwen;Parthasarathy,Varsha;Kwatra,ShawnG;Glass2nd,DonaldA
• 通讯作者: Glass2nd,DonaldA