The Roles of a Novel Microprotein in Wound Healing and Cancer

新型微生物蛋白在伤口愈合和癌症中的作用

• 批准号: 10682567
• 负责人: Thomas Farid Martinez
• 金额: $ 17.2万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10682567
• 关键词: 3-Dimensional; Advisory Committees; Affect; Amino Acids; Binding; Bioinformatics; Biology; Breast Cancer Cell; Breast Cancer cell line; CRISPR screen; Cancer Biology; Cell Proliferation; Cells; Chromatin; Chromatin Remodeling Factor; Code; Complex; Core Facility; Data; Data Set; Deacetylase; Dependence; Development; Developmental Biology; Disease; Educational workshop; Environment; Enzymes; Epidermis; Epigenetic Process; Fostering; Foundations; Gene Expression; Genes; Genetic Transcription; Genomics; Goals; HDAC1 gene; Histone Deacetylation; Histones; Homeostasis; Human; Human Cell Line; In Vitro; K-Series Research Career Programs; Knockout Mice; Knowledge; Malignant Neoplasms; Malignant neoplasm of prostate; Mammals; Mediating; Mentors; Mentorship; MicroRNAs; Modeling; Modification; Molecular; NuRD complex; Open Reading Frames; Pathway interactions; Peptides; Phenotype; Process; Proliferating; Proteins; Proteomics; RNA; Regulation; Research; Role; Running; Signal Pathway; Skin; Skin wound healing; Techniques; Testing; Thick; Tissues; Training; Translating; Tumor Promotion; Undifferentiated; Untranslated RNA; cancer cell; cancer therapy; career; cell motility; chromatin remodeling; data integration; differential expression; epigenetic regulation; experience; experimental study; in vivo; innovation; keratinocyte; knock-down; malignant breast neoplasm; migration; mouse model; new therapeutic target; novel; programs; prostate cancer cell; prostate cancer cell line; protein function; ribosome profiling; skin wound; transcriptome; transcriptome sequencing; tumor; tumor progression; tumorigenesis; wound; wound closure; wound healing

Project Summary The pathways that regulate processes necessary for wound healing, such as proliferation and migration, are often co-opted by tumors, leading to their description as wounds that do not heal. Therefore, identifying novel genes involved in wound healing and understanding how they are dysregulated in cancer may provide new targets for therapeutic efforts. We recently discovered thousands of small open reading frames that encode proteins <100 amino acids, dubbed microproteins. Among these was a 10 kDa microprotein encoded on the lncRNA Terminal Differentiation-Induced non-coding RNA (TINCR), which is a critical inducer of terminal differentiation in the epidermis and regulator of cancer cell proliferation and migration. TINCR microprotein (TINCR-MP) is highly conserved across mammals, strongly suggesting that it is functional. Compelling preliminary data demonstrate that in human skin models TINCR-MP downregulates signaling pathways involved in wound healing, and that it interacts with histone modifying enzymes that have functions in differentiation and cancer. Furthermore, while TINCR-MP is expressed during epidermal differentiation, it is unnecessary for differentiation, making its function separate from the differentiation promoting activity of TINCR RNA. The central hypothesis is that TINCR-MP acts as a brake for proliferation and migration during wound healing through alterations to the epigenetic landscape, and that this function is hijacked in cancer cells to promote tumor progression. The goals of this proposal are thus: 1) to determine the role of TINCR-MP in cutaneous wound repair processes, and establish whether TINCR-MP effects on wound healing are driven by epigenetic modifications, 2) to determine the extent to which TINCR-MP regulates proliferation and migration in prostate and breast cancer cell lines, and establish its effects on epigenetic and subsequent gene expression changes, and 3) to identify additional microproteins regulated during differentiation that affect cancer proliferation. Successful completion of these aims will reveal an important microprotein that functions in wound healing and tumor progression, as well as new microproteins to investigate. The candidate, Dr. Thomas Martinez, plans to develop an independent research program focused on characterizing microproteins that function in both differentiation and cancer. The opportunities offered by this Career Development Award will allow Dr. Martinez to deepen his knowledge of developmental biology, epigenetics, and cancer biology. He will also gain experience utilizing 3D ex vivo skin models, in vivo mouse models, and techniques for analyzing the epigenetic landscape. Dr. Martinez will conduct these studies under the mentorship of Dr. Alan Saghatelian, expert on peptide biology, as well as co-mentors Dr. Diana Hargreaves and Dr. George Sen, experts on chromatin remodeling complexes in disease and epidermal homeostasis, respectively. Dr. Martinez’s Advisory Committee will also help foster his scientific and academic career goals. The Salk Institute provides an ideal environment with expertly run core facilities and ample seminars and workshops to prepare trainees for independent academic careers.

项目摘要 调节伤口愈合所必需的过程（如增殖和迁移）的途径是 通常被肿瘤吸收，导致它们被描述为无法愈合的伤口。因此，识别新 参与伤口愈合的基因以及了解它们在癌症中是如何失调的可能会提供新的 治疗努力的目标。我们最近发现了数千个小的开放阅读框架， 小于100个氨基酸的蛋白质，称为微蛋白。其中有一个10 kDa的微蛋白编码的， lncRNA末端分化诱导的非编码RNA（TINCR），其是末端分化的关键诱导剂。 在表皮中的分化和癌细胞增殖和迁移的调节剂。TINCR微蛋白 （TINCR-MP）在哺乳动物中高度保守，强烈表明它是功能性的。引人注目 初步数据表明，在人类皮肤模型中，TINCR-MP下调了涉及 在伤口愈合中，它与组蛋白修饰酶相互作用，组蛋白修饰酶在分化和 癌此外，虽然TINCR-MP在表皮分化期间表达，但它对于表皮细胞的分化是不必要的。 TINCR RNA的分化促进活性，使其功能与TINCR RNA的分化促进活性分开。中央 假设TINCR-MP在伤口愈合过程中作为增殖和迁移的制动器， 表观遗传景观的改变，并且这种功能在癌细胞中被劫持以促进肿瘤生长。 进展因此，本提案的目标是：1）确定TINCR-MP在皮肤创伤中的作用 修复过程，并确定TINCR-MP对伤口愈合的影响是否由表观遗传驱动 2）确定TINCR-MP调节前列腺中增殖和迁移的程度 和乳腺癌细胞系，并建立其对表观遗传和随后的基因表达变化的影响， 和3）鉴定在分化过程中调节的影响癌增殖的其它微蛋白。 这些目标的成功完成将揭示一种重要的微蛋白，它在伤口愈合中起作用， 肿瘤进展，以及新的微蛋白研究。候选人托马斯马丁内斯博士计划 开发一个独立的研究计划，重点是表征微蛋白，在这两个功能 分化和癌症。这个职业发展奖提供的机会将使马丁内斯博士 以加深他对发育生物学、表观遗传学和癌症生物学的了解。他也将获得经验 利用3D离体皮肤模型、体内小鼠模型和用于分析表观遗传景观的技术。 博士Martinez将在肽生物学专家Alan Saghatelian博士的指导下进行这些研究， 以及共同导师戴安娜·哈格里夫斯博士和乔治森博士，染色质重塑复合物专家 在疾病和表皮稳态中的作用。马丁内斯博士的咨询委员会也将帮助促进他的 科学和学术职业目标。索尔克研究所提供了一个理想的环境， 设施和充足的研讨会和讲习班，为学员的独立学术生涯做好准备。

项目成果

Integrated workflow for discovery of microprotein-coding small open reading frames.

• DOI: 10.1016/j.xpro.2023.102649
• 发表时间: 2023-12-15
• 期刊: STAR PROTOCOLS
• 作者: Cao, Kevin;Heydary, Yasamin Hajy;Tong, Gregory;Martinez, Thomas Farid
• 通讯作者: Martinez, Thomas Farid