Human-Enzyme Mediated, Systemic Depletion of Cystine for Cancer Treatment.

人类酶介导的胱氨酸系统消耗用于癌症治疗。

基本信息

批准号：
8785755
负责人：
John DiGiovanni
金额：
$ 58.04万
依托单位：
UNIVERSITY OF TEXAS AT AUSTIN
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-08-01 至 2019-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8785755
关键词：
Ablation Acute Lymphocytic Leukemia Alanine Allografting American Amino Acid Transport System L Amino Acid Transporter Amino Acids Androgens Animal Model Anti-Inflammatory Agents Anti-inflammatory Antineoplastic Agents Antioxidants Apoptosis Autophagocytosis Biochemical Biodistribution Biological Markers Body Weight decreased Brain Neoplasms Breast Carcinoma Cancer Cell Growth Cancer Etiology Cancer Model Cell Cycle Arrest Cell Line Cell membrane Cells Cessation of life Chemistry Clinical Combined Modality Therapy Cyst Cystathionine Cysteine Cystine Data Defect Dependence Development Disease Dose Energy Metabolism Engineering Enzyme Kinetics Enzymes Equilibrium Essential Amino Acids Extracellular Matrix Extracellular Space Genetic Glioblastoma Glioma Glutamates Goals Growth Homeostasis Human Human Engineering Immunosuppression In Vitro Lead Long-Term Effects Lung Lyase Malignant Epithelial Cell Malignant Neoplasms Malignant neoplasm of prostate Mammals Mediating Metabolic Methionine Modality Modeling Mus Mutation Names Neoplasms Non-Essential Amino Acid Non-Malignant Normal tissue morphology Oxidation-Reduction Oxidative Stress Pancreatic carcinoma Pathway interactions Pegaspargase Pharmaceutical Preparations Pharmacodynamics Phenotype Predisposition Process Production Prostate Prostate carcinoma Prostatic Neoplasms Protein Biosynthesis Proteins Quality of life Reactive Oxygen Species Relative (related person)Research Resistance Route Serine Serum Signal Pathway Signal Transduction Staging Structure Sulfasalazine Sulfur System Therapeutic Tissues Toxic effect Tumor Tissue Validation Work Xenograft procedure antiporter base bioprocess cancer cell cancer stem cell cancer therapy cancer type chemotherapy clinically relevant deprivation design drug efficacy experience extracellular high throughput screening human FRAP1 protein improved in vivo lung small cell carcinoma meetings men metabolomics mortality mouse model neoplastic cell novel novel therapeutic intervention pre-clinical public health relevance research study response small molecule standard of care success therapeutic enzyme therapeutic target tool treatment effect tumor tumor growth uptake

项目摘要

DESCRIPTION (provided by applicant): Extensive studies have revealed that in contrast to normal tissue, prostate carcinomas (PCa) and other high mortality cancers including glioblastomas, and small cell lung carcinomas are critically dependent on the uptake of the non-essential amino acid L-cysteine (L-Cys) and its oxidized form L-cystine (CSSC) for proliferation and survival (due to increased requirements for anti-oxidants and in some tumors because they display defects in endogenous L-Cys synthesis). We hypothesize that enzyme-mediated systemic depletion of the serum L-Cys/CSSC pool (using an engineered human enzyme, and thus likely well tolerated) constitutes a powerful and completely novel cancer therapeutic approach for PCa as a monotherapy and as an agent that is mechanistically synergistic with existing chemotherapeutic modalities. This hypothesis is supported by strong preliminary data demonstrating that the administration (every 4 days) of a prototypical engineered human L- Cys/CSSC depleting enzyme in an allograft model of PCa: (i) results in near complete depletion of the serum L-Cys/CSSC pool; (ii) mediates sustained and complete cessation of tumor growth with cancer cells experiencing cell cycle arrest accompanied by increased oxidative stress and autophagy and (iii) importantly treatment for over a month was very well tolerated with no weight loss or gross toxicities. To build on these preliminary observations we present a detailed research plan that is aimed at delivering a clinically relevant enzyme therapeutic candidate and providing the validation by in vitro and in allograft/xenograft PCa models that will eventually hel support late stage preclinical development and an IND application. While L-Cys/CSSC depletion may be applicable to the treatment of many tumors we elected to focus on PCa first because many human PCA tumors are known to experience defects in L-Cys/CSSC homeostasis and second because effective non-toxic treatments are needed as PCa is the second leading cause of cancer death in American men. Reaching these goals will require the completion of work under 3 Specific Aims: Sp. Aim 1 will focus on the engineering of a candidate human enzyme for L-Cys/CSSC depletion that displays optimal enzyme kinetics, substrate selectivity, stability to deactivation and aggregation and facile production by bioprocessing. The PK and PD of the optimized enzymes as well as any toxic effects upon repeated administration will be evaluated in detail. In Sp. Aim 2 we will seek to understand the global mechanistic/metabolic consequences of L-Cys/CSSC depletion on PCa cells, what biomarkers may predict susceptibility/resistance and to determine which combinations with standard of care chemotherapeutics may provide synergistic/additive effects. Finally, in Sp. Aim 3 we will evaluate the optimized enzyme in clinically relevant orthotopic PCa animal models that represent diverse phenotypes (i.e. p53 status, androgen dependence/independence) and mouse orthotopic PCa allografts such that the efficacy of the drug can be studied in models closely mimicking the actual disease.

描述（由申请人提供）：广泛的研究表明，与正常组织，前列腺癌（PCA）和其他高死亡率癌症相反，包括胶质母细胞瘤以及小细胞肺癌在关键上取决于非氨基酸L-Cysteine（L-Cys）和氧化氨基（L-Cys）及其氧化（L-Cysif（L-Cysif）（l-Cysif（L-Cysif）（cysif）（l-Cysif（L-Cysif））（cysif（L-Cys））的氧化（L-Cysif（L-Cysif）（Cy））对抗氧化剂和某些肿瘤的需求增加，因为它们在内源性L-CYS合成中表现出缺陷）。我们假设酶介导的血清L-CYS/CSSC池的全身性耗竭（使用工程的人酶，因此可能耐受性良好）构成了PCA作为单一疗法的强大且完全新颖的癌症治疗方法，并且是一种机械性化学疗法的药物，具有现有的化学疗法模式。该假设得到了强大的初步数据的支持，表明在PCA的同种异体移植模型中，典型的人类L-CYS/CSSC耗尽酶的给药（每4天）几乎完全消耗了Serum L-Cys/cssc池；（ii）介导肿瘤生长的持续和完全停止，癌细胞经历细胞周期停滞，伴随着氧化应激和自噬的增加，并且（iii）重要的是，一个月以上的治疗非常耐受，没有体重减轻或严重的毒性。为了基于这些初步观察，我们提出了一项详细的研究计划，旨在提供临床相关的酶治疗候选候选者，并通过体外和同种异体移植/Xenograft PCA模型提供验证，该模型最终将支持后期的临时开发和IND应用。尽管L-CYS/CSSC耗竭可能适用于对许多肿瘤的治疗，但我们首先选择专注于PCA，因为已知许多人PCA肿瘤在L-CYS/CSSC体内稳态中经历缺陷，第二个是因为PCA是美国男性癌症死亡的第二个主要原因，因此需要有效的无毒治疗。达到这些目标将需要以3个特定目标完成工作：sp。 AIM 1将重点介绍候选人酶的L-CYS/CSSC耗竭，该酶显示出最佳的酶动力学，底物选择性，稳定性通过生物处理，停用，聚集和便利产生。优化酶的PK和PD以及对重复给药的任何有毒作用将进行详细评估。在sp。 AIM 2我们将寻求了解L-CYS/CSSC耗竭对PCA细胞的全球机械/代谢后果，哪些生物标志物可以预测易感性/耐药性，并确定哪种与护理标准化学治疗剂的组合可能会提供协同/添加剂。最后，在sp。 AIM 3我们将评估代表不同表型（即p53状态，雄激素依赖/独立性）和小鼠原位PCA同种异体移植物的临床相关的原始PCA动物模型中优化的酶，以使该药物的功效在模型中可以仔细模仿实际疾病的模型中进行研究。