Development of Microbial-Based Therapies to Suppress Macropinocytosis in Kras-Driven Cancers

开发基于微生物的疗法来抑制 Kras 驱动的癌症中的巨胞饮作用

• 批准号: 10502177
• 负责人: EDWIN MANUEL
• 金额: $ 40.26万
• 依托单位: BECKMAN RESEARCH INSTITUTE/CITY OF HOPE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10502177
• 关键词: Affinity; Anabolism; Antimetabolites; Attenuated; Automobile Driving; Autophagocytosis; Binding; Bladder; Breast; Catabolic Process; Cell Line; Cell Proliferation; Cell surface; Clinical; Colon; Colorectal Cancer; Culture Media; Development; Disease; Drug resistance; Endocytosis; Engineering; Enzymes; Extracellular Matrix; Fluorouracil; Gene Expression; Genes; Glucosamine; Glucuronic Acids; Glycosides; Goals; Growth; Growth Factor; Heparan Sulfate Proteoglycan; Heparin Lyase; Heparitin Sulfate; Homeostasis; Investigational Drugs; KRAS oncogenesis; Lesion; Lung; Lysosomes; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of pancreas; Malignant neoplasm of prostate; Malignant neoplasm of urinary bladder; Manuals; Measures; Mediating; Mediator of activation protein; Metabolic; Metabolism; Methods; Modification; Monomeric GTP-Binding Proteins; National Cancer Institute; Natural regeneration; Neoplasm Metastasis; Nutrient; Oncogenic; Outcome; Pancreatic Ductal Adenocarcinoma; Pathway interactions; Patients; Plasmids; Process; Production; Prognosis; Prostate; Proteins; Recombinants; Recurrence; Research; Resistance; Salmonella typhimurium; Signal Transduction; Signaling Molecule; Source; Stress; Structure; Testing; Therapeutic; Therapeutic Agents; Therapeutic Use Study; Tumor Tissue; Vertebral column; Work; Xenograft Model; base; cancer therapy; clinical translation; clinically relevant; conventional therapy; cost; cytokine; depolymerization; design and construction; experience; extracellular; gemcitabine; heparanase; heparinase III; in vivo; ineffective therapies; inhibitor; macromolecule; malignant breast neoplasm; microbial; mortality; mouse model; mutant; neoplastic cell; neovascularization; novel; novel strategies; pancreatic ductal adenocarcinoma cell; pancreatic ductal adenocarcinoma model; patient derived xenograft model; promoter; receptor; response; standard of care; syndecan; targeted agent; targeted treatment; therapeutic target; therapy resistant; tumor; tumor growth; tumor metabolism; tumor microenvironment; tumor progression; tumorigenic; vector

PROJECT SUMMARY Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive disease with dismal prognosis. A near-universal oncogenic driver of PDAC is the constitutive activation of the small GTPase protein Kras, which induces multiple downstream signaling cascades that together facilitate rapid cell proliferation, metastasis and therapeutic resistance. To surmount the high energy demands of these activities, Kras also triggers metabolic adaptations to promote nutrient scavenging from extracellular sources, such as through macropinocytosis. Macropinocytosis is a process by which extracellular material is non-specifically engulfed and then degraded in lysosomes to produce end-products utilized by tumor cells for biosynthesis. This process essentially confers resistance to a myriad of anabolic inhibitors. Syndecan-1 is a heparan sulfate proteoglycan (HSPG) upregulated on the surface of cells that serves as the key mediator of macropinocytosis in PDAC and other Kras-driven cancers that includes bladder, lung, prostate, colon and breast. In addition to mediating macromolecular transport, HSPGs can be found in the tumor extracellular matrix (ECM) binding to and regulating the interaction of numerous signaling molecules (e.g. growth factors and cytokines) with their cognate receptors. The pro-tumorigenic activities of HSPGs are exquisitely regulated by enzymatic modification of their heparan sulfate (HS) moieties. Mammalian heparanases employ hydrolytic cleavage of the beta-(1,4)-glycosidic bond between glucuronic acid and glucosamine to promote the release of growth factors and enzymes involved in ECM remodeling, invasion and metastasis. In contrast to mammalian heparanases, bacterial heparinase III (HepIII) depolymerizes HSPGs through a unique beta-elimination mechanism that cleaves at the alpha-(1,4)- glycosidic bond. Various studies have confirmed HepIII modification of HSPGs suppresses neovascularization, macropinocytosis, tumor growth and metastasis. However, the inability to restrict HepIII activity to tumor tissue has long prohibited its use as a therapeutic agent. Using attenuated, tumor-targeting Salmonella typhimurium (ST) vectors, we have developed the first recombinant ST expressing functional HepIII (ST-HepIII) through a tightly regulated, inducible promoter. We have confirmed the ability of ST-HepIII to suppress high-affinity HS interactions, macropinocytosis, and growth of Kras-mutant tumors. In this application, we will: 1) Determine the impact of ST-HepIII treatment on metabolite availability and metabolic-associated gene pathways in vivo; 2) Determine anti-tumor efficacy of anabolic inhibitors in combination with ST-HepIII; and 3) Develop and characterize recombinant STs expressing HepIII under tumor-inducible promoters for greater clinical feasibility. Completing these aims will allow us to develop a novel class of tumor-targeting agents capable of suppressing a metabolic process essential to the survival of PDAC and other Kras-driven cancers. Our agents may be used to counteract acquired resistance to standard-of-care therapies that target cooperative anabolic processes and positively impact survival for patients with difficult-to-treat cancers.

胰腺导管腺癌（PDAC）是一种高度侵袭性的疾病， 预后PDAC的一个几乎普遍的致癌驱动因素是小GT3蛋白的组成性激活， Kras，它诱导多个下游信号级联，共同促进快速细胞增殖， 转移和治疗抗性。为了克服这些活动的高能量需求，Kras还 触发代谢适应，以促进从细胞外来源清除营养素，例如通过 巨胞饮作用巨胞饮作用是细胞外物质被非特异性吞噬的过程， 然后在溶酶体中降解以产生终产物，所述终产物被肿瘤细胞用于生物合成。这个过程 基本上赋予了对无数合成代谢抑制剂的抗性。Syndecan-1是一种硫酸乙酰肝素蛋白聚糖 HSPG在细胞表面上调，作为PDAC中巨胞饮的关键介质， 其他Kras驱动的癌症，包括膀胱，肺，前列腺，结肠和乳腺癌。除了调解 在大分子转运中，HSPGs可以在肿瘤细胞外基质（ECM）中发现， 许多信号分子（如生长因子和细胞因子）与其同源受体的相互作用。 HSPGs的促肿瘤活性通过其乙酰肝素的酶促修饰而受到精细调控 硫酸酯（HS）部分。哺乳动物类肝素酶利用β-（1，4）-糖苷键的水解裂解 葡萄糖醛酸和葡萄糖胺之间的相互作用，以促进生长因子和酶的释放， ECM重塑、侵袭和转移。与哺乳动物肝素酶相反，细菌肝素酶III （HepIII）通过独特的β-消除机制解聚HSPG，该机制在α-（1，4）- 糖苷键各种研究已经证实HSPG的HepIII修饰抑制新血管形成， 巨胞饮作用、肿瘤生长和转移。然而，不能将HepIII活性限制在肿瘤组织中， 长期以来一直禁止将其用作治疗剂。使用减毒的、针对肿瘤的鼠伤寒沙门氏菌 (ST)载体，我们已经开发了第一个重组ST表达功能性HepIII（ST-HepIII），通过 严格调控的诱导型启动子。我们已经证实了ST-HepIII抑制高亲和力HS的能力， 相互作用、巨胞饮作用和Kras突变型肿瘤的生长。在本申请中，我们将：1）确定 ST-HepIII治疗对体内代谢物利用率和代谢相关基因途径影响; 2） 确定合成代谢抑制剂与ST-HepIII组合的抗肿瘤功效;以及 表征在肿瘤诱导型启动子下表达HepIII的重组ST，以获得更大的临床可行性。 完成这些目标将使我们能够开发出一类新的肿瘤靶向药物，能够抑制 这是PDAC和其他Kras驱动的癌症生存所必需的代谢过程。我们的代理人可能会被用于 抵消对靶向协同合成代谢过程的标准治疗的获得性耐药性， 对难治性癌症患者的生存率有积极影响。