DISSECTING AND TARGETING THE ROLE OF GALNT14 IN HIGH-RISK OSTEOSARCOMA

剖析和瞄准 GALNT14 在高风险骨肉瘤中的作用

• 批准号: 10761850
• 负责人: Jennifer J Kohler
• 金额: $ 19.37万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-13 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10761850
• 关键词: Address; Adjuvant Chemotherapy; Adolescent; Adult; Alternative Therapies; Antigens; Azides; Biological Assay; Biology; Bone neoplasms; Breast; CRISPR/Cas technology; Categories; Cell Communication; Cell Culture Techniques; Cell Line; Cells; Chemicals; Chemoresistance; Chemotherapy-Oncologic Procedure; Childhood Osteosarcoma; Cisplatin; Clinical Trials; Cultured Cells; Databases; Diagnosis; Disease Progression; Disease Resistance; Disease-Free Survival; Dose; Doxorubicin; Drug Kinetics; Enzyme-Linked Immunosorbent Assay; Enzymes; Evaluation; Excision; Exhibits; Family; Future; Genes; Glycobiology; Goals; Immune system; Immunotherapeutic agent; In Vitro; Incidence; Inferior; Informatics; Institution; Knock-out; Libraries; Link; Lung; Malignant Neoplasms; Mass Spectrum Analysis; Measures; Methods; Methotrexate; Mission; Modality; Modeling; Molecular; Molecular Target; Necrosis; Neoadjuvant Therapy; Neoplasm Metastasis; Ontology; Operative Surgical Procedures; Outcome; Patient-Focused Outcomes; Patients; Pediatric Neoplasm; Permeability; Post-Translational Protein Processing; Primary Neoplasm; Prognosis; Prognostic Factor; Prostate; Protein Isoforms; Proteins; Public Health; Rapid screening; Recombinants; Recurrent disease; Refractory; Refractory Disease; Regimen; Relapse; Research; Resected; Resistance; Role; Serine; Signal Transduction; Survival Rate; Testing; Therapeutic; Threonine; Toxic effect; United States National Institutes of Health; chemotherapy; childhood sarcoma; clinical development; effective therapy; experience; functional genomics; gain of function; glycosylation; high risk; high throughput screening; improved; in vitro Model; in vivo; inhibitor; insight; lead candidate; loss of function; new therapeutic target; novel; osteosarcoma; overexpression; patient derived xenograft model; predicting response; protein transport; resistance mechanism; response; sarcoma; small molecule; small molecule inhibitor; standard of care; sugar; synergism; targeted treatment; therapeutic candidate; therapeutic target; therapy resistant; transcriptomics; treatment response; treatment strategy; tumor; tumor growth; tumor progression

Osteosarcoma (OS) is the most common bone tumor in children and adolescents. Regimens of neoadjuvant doxorubicin, cisplatin, and high dose methotrexate have been the standard of care since the 1970s, but no additional chemo or targeted therapies have added significant survival benefits, leaving minimal options for relapsed and resistant disease. These patients have an extremely poor prognosis, with long term outcomes of less than 20%. Therefore, a better understanding of chemoresistance mechanisms is essential towards making our current therapies more effective. Through comprehensive transcriptomic analysis of institutional chemoresistant patient-derived xenograft (PDX) models and the publicly available Therapeutically Applicable Research to Generate Effective Treatments (TARGET) OS database, we identified significant elevation of glycosylation genes in patients with poor response to chemotherapy. Glycosylation is a form of protein post- translational modification that plays a role in protein trafficking, function, and stability. It also provides substrates for cell-cell interactions, antigens for immune system surveillance, and often plays a role in cell signaling. O-type glycosylation, the addition of sugar moieties to serine or threonine residues to proteins, begins with the family of N-acetylgalactosyltransferase enzymes (GALNTs). Our analysis identified overexpression of GALNT14 in poor responding (<90% necrosis) OS tumors and we provide evidence that expression of GALNT14 has significant inverse correlation with overall survival and event-free survival in OS. GALNT14 has been shown to promote chemotherapy resistance and metastasis in adult cancers, such as of the breast, prostate, and lung. Our hypothesis is GALNT14 has a significant impact on disease progression in pediatric OS by promoting chemotherapy resistance and is a novel, viable therapeutic target for small molecule inhibition. We propose two complementary, yet independent aims to address this hypothesis. The proposal will synergize the expertise of Dr. Yustein, in sarcoma biology and modeling, and Dr. Kohler in glycobiology to successfully accomplish the goals. AIM1: Role of GALNT14 in promoting chemoresistance in osteosarcoma models in vitro and in vivo. Using both commercially available and PDX-derived chemoresistant cell lines we are developing both gain and loss-of-function GALNT14 models through overexpression and CRISPR/Cas9 knockout approaches. Using these models, we will evaluate in vitro and in vivo effects on OS sensitivity to chemotherapy and effects on tumor growth and progression. AIM2. Identification and evaluation of small molecule inhibitors of GALNT14. In this aim, we will use high-throughput screening to identify small molecules that inhibit the activity of purified, recombinant GALNT14. Further, we will determine which of these molecules can inhibit GALNT14 in cell culture. GALNT14 inhibitors identified in this aim have the potential to serve as lead candidates for clinical development and may also be used to investigate the mechanistic involvement of GALNT14 in OS. Overall, our studies will lead to the identification of critical candidate therapeutic modalities for the treatment of high-risk OS.

骨肉瘤（OS）是儿童和青少年最常见的骨肿瘤。新辅助方案 阿霉素，顺铂和高剂量甲氨蝶呤一直是1970年代以来的护理标准，但没有 其他化学疗法或靶向疗法增加了显着的生存益处，为 复发和抗性疾病。这些患者的预后非常差，长期结局 小于20％。因此，对化学抗性机制的更好理解对于制造 我们目前的疗法更有效。通过制度的全面转录组分析 化学耐药的患者衍生异种移植（PDX）模型和公开可用的治疗疗法 研究生成有效治疗（目标）OS数据库的研究，我们确定了显着的高度升高 对化学疗法反应不佳的患者的糖基化基因。糖基化是蛋白质后的一种形式 转化修饰在蛋白质运输，功能和稳定性中起作用。它还提供基材 对于细胞 - 细胞相互作用，用于免疫系统监测的抗原，并且通常在细胞信号中起作用。 O型 糖基化，将糖部分添加到丝氨酸或苏氨酸残基到蛋白质中，始于 N-乙酰乳糖基转移酶（GALNTS）。我们的分析确定了差的Galnt14的过表达 反应（<90％坏死）OS肿瘤，我们提供的证据表明GALNT14的表达显着 与OS中的总生存期和无事件生存期的逆相关性。 GALNT14已被证明可以促进 成年癌症（例如乳房，前列腺和肺部）的化学疗法耐药性和转移。我们的 假设是GALNT14通过促进小儿OS的疾病进展有重大影响 化学疗法耐药性，是一个新型，可行的小分子抑制靶标。我们提出了两个 补充，但独立的旨在解决这一假设。该提案将协同作用 Yustein博士，专业的肉瘤生物学和建模，Kohler博士在糖生物学领域成功完成 目标。 AIM1：GALNT14在体外和在体外促进骨肉瘤模型中促进化学上的作用 体内。使用市售和PDX衍生的化学剂细胞系，我们都在开发两种增益 通过过表达和CRISPR/CAS9敲除方法，功能丧失的GALNT14模型。使用 这些模型，我们将评估体外和体内对OS对化学疗法敏感性的影响以及对肿瘤的影响 生长和发展。 AIM2。 GALNT14的小分子抑制剂的鉴定和评估。在 这个目的，我们将使用高通量筛选来识别抑制纯化活性的小分子 重组GALNT14。此外，我们将确定哪些分子可以抑制细胞培养中的GALNT14。 该目标中确定的GALNT14抑制剂有可能作为临床开发的主要候选者 并且还可以用于研究GALNT14在OS中的机械参与。总的来说，我们的研究将 导致鉴定关键的候选治疗方法用于治疗高危OS。