KISS1: Defining Mechanisms for Antimetastatic Therapy

KISS1：抗转移治疗的定义机制

• 批准号: 8676690
• 负责人: Danny R. Welch
• 金额: $ 28.12万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8676690
• 关键词: Acute; Address; Agonist; Binding; Biological; Blast Cell; Cells; Cessation of life; Chronic; Clinic; Clinical; Clinical Management; Coculture Techniques; Complementary DNA; Data; Development; Disease; Disseminated Malignant Neoplasm; Ectopic Expression; Engineering; Feedback; Fibroblasts; Fluorescence Microscopy; Future; G Protein-Coupled Receptor 54; Growth; Human; In Situ; In Vitro; KISS1 gene; KISS1R gene; Length; Ligands; Lung; Malignant Neoplasms; Melanoma Cell; Metastasis Suppression; Microscopic; Molecular Target; Morbidity - disease rate; Mutate; Neoplasm Metastasis; Normal Cell; Paracrine Communication; Peptides; Physiological; Primary Neoplasm; Process; Production; Prohormone Convertase; Proliferating; Publications; Quality of life; Signal Transduction; Site; Site-Directed Mutagenesis; Skin; Stromal Cells; Symptoms; Testing; Tetanus Helper Peptide; Tetracyclines; Therapeutic; Time; Tissues; Tumorigenicity; Work; autocrine; base; cancer therapy; cellular targeting; expression vector; health economics; human KISS1 protein; insight; kisspeptin; mortality; mutant; neoplastic cell; novel; novel strategies; paracrine; prevent; promoter; research study; response; tumor

DESCRIPTION (provided by applicant): Since the majority of cancer deaths and morbidity are due to metastases, major improvements in survival and quality of life will occur when metastasis are prevented or more effectively treated. Determining whether the KISS1 metastasis suppressor can maintain disseminated tumor cells in a non- proliferating, dormant state may provide a novel approach to cancer therapy. BACKGROUND: Re-expression of KISS1 blocks the ability of melanoma cells to colonize multiple ectopic sites while still completing antecedent steps of the metastatic cascade. KISS1 is processed into numerous peptides, termed kisspeptins. Some kisspeptins bind to and stimulate a G-protein coupled receptor, GPR54; however, metastasis suppression does not appear to require tumor cell GPR54 expression. HYPOTHESIS #1: KISS1 re- expression will halt further growth of established (micro)metastases. Specific Aim 1: Using Tet-inducible expression vectors in GFP-expressing melanoma cells seeding lung, KISS1 expression will be induced or turned off when lung foci are different sizes. Further growth, regression or induced dormancy will be assessed by fluorescence microscopy. Implications: If KISS1 could halt progression or reverse established metastases, its utility for treatment of human cancer would increase significantly. HYPOTHESIS #2: Selected kisspeptins are responsible for metastasis suppression. Specific Aim 2: Preliminary data show that KISS1 -> kisspeptins processing occurs outside the cell, but which kisspeptin(s) suppresses metastasis has not been determined. Using site-directed mutagenesis, disruption of KISS1->kisspeptin processing sites (R-R or R-K) in KISS1 and ectopic expression of the processing mutants will be done followed by assessment of kisspeptin-induced signaling thru GPR54 and metastatic potential. Implications: Identifying which kisspeptin(s) are responsible for metastasis suppression will focus future agonist development. HYPOTHESIS #3: KISS1 suppresses metastasis via paracrine signaling with stromal cells. Specific Aim 3: Preliminary data show that tumor cells suppressed by re-expression of KISS1 do not express GPR54, suggesting that KISS1 production by tumor cells acts via intermediary cells (i.e., paracrine). The initial working hypothesis is that stromal fibroblasts are the intermediary cell because they express GPR54 in vitro. Using RTQ and IHC, we will determine which stromal cells express GPR54 in situ. 2D and 3D co-culture of parental and KISS1-expressing cells with fibro- blasts from different tissues will assess whether differential response of tissue stroma to KISS1 returns growth promoting or growth inhibitory signals to tumor cells. Implications: These experiments ultimately test what the actual target of KISS1 is with regard to metastasis suppression. Utilization in the clinic will vary depending upon whether one is targeting tumor cells or normal cells. Data from the proposed experiments will determine the molecular and cellular target(s) of specific kisspeptin(s) and aspects of the timing of exposure required to suppress metastasis. Those data will be crucial if KISS1 is to be developed as an anti-metastatic therapy.

描述（由申请人提供）：由于大多数癌症死亡和发病都是由转移引起的，因此当预防转移或更有效地治疗转移时，生存率和生活质量将出现重大改善。确定KISS1转移抑制因子是否可以将播散的肿瘤细胞维持在非增殖、休眠状态可能为癌症治疗提供一种新方法。背景：KISS1 的重新表达会阻断黑色素瘤细胞定植多个异位位点的能力，同时仍能完成转移级联的先前步骤。 KISS1 被加工成多种肽，称为 Kisspeptins。一些 Kisspeptins 结合并刺激 G 蛋白偶联受体 GPR54；然而，转移抑制似乎并不需要肿瘤细胞表达 GPR54。假设#1：KISS1 重新表达将阻止已建立的（微）转移瘤的进一步生长。具体目标1：在接种肺部的表达GFP的黑色素瘤细胞中使用Tet诱导型表达载体，当肺病灶大小不同时，KISS1表达将被诱导或关闭。将通过荧光显微镜评估进一步的生长、消退或诱导的休眠。意义：如果 KISS1 能够阻止进展或逆转已形成的转移，那么其治疗人类癌症的效用将显着增加。假设#2：选定的 Kisspeptins 负责抑制转移。具体目标 2：初步数据显示 KISS1 -> Kisspeptins 加工发生在细胞外，但哪种 Kisspeptin 抑制转移尚未确定。使用定点诱变，破坏 KISS1 中的 KISS1->Kisspeptin 加工位点（R-R 或 R-K）以及加工突变体的异位表达，然后评估 Kisspeptin 通过 GPR54 诱导的信号传导和转移潜力。意义：确定哪些 Kisspeptin 负责抑制转移将集中未来激动剂的开发。假设#3：KISS1 通过与基质细胞的旁分泌信号传导抑制转移。具体目标 3：初步数据显示，被 KISS1 重新表达抑制的肿瘤细胞不表达 GPR54，这表明肿瘤细胞产生 KISS1 通过中间细胞（即旁分泌）发挥作用。最初的工作假设是基质成纤维细胞是中间细胞，因为它们在体外表达 GPR54。使用 RTQ 和 IHC，我们将确定哪些基质细胞原位表达 GPR54。亲代细胞和表达 KISS1 的细胞与来自不同组织的成纤维细胞的 2D 和 3D 共培养将评估组织基质对 KISS1 的差异反应是否向肿瘤细胞返回生长促进或生长抑制信号。意义：这些实验最终测试了 KISS1 在抑制转移方面的实际目标是什么。临床上的利用将根据是针对肿瘤细胞还是正常细胞而有所不同。拟议实验的数据将确定特定 Kisspeptin 的分子和细胞靶点以及抑制转移所需的暴露时间。如果 KISS1 被开发为抗转移疗法，这些数据将至关重要。

项目成果

Tumor Cell Invasion-Not All Barriers Are Created Equal.

• DOI: 10.1158/0008-5472.can-16-0550
• 发表时间: 2016-04-01
• 期刊: Cancer research
• 影响因子: 11.2
• 作者: Welch DR

Unraveling the 'TGF-β paradox' one metastamir at a time.

• DOI: 10.1186/bcr3383
• 发表时间: 2013-02-27
• 期刊: Breast cancer research : BCR
• 作者: Welch DR;Hurst DR
• 通讯作者: Hurst DR