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: 10652633
• 负责人: EDWIN MANUEL
• 金额: $ 39.45万
• 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/10652633
• 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; Goals; Growth; Growth Factor; Heparan Sulfate Proteoglycan; Heparin Lyase; Heparitin Sulfate; Homeostasis; Invaded; 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; Metabolic; Metabolism; Methods; Modification; Monomeric GTP-Binding Proteins; National Cancer Institute; Neoplasm Metastasis; Nutrient; Oncogenic; Outcome; Pancreatic Ductal Adenocarcinoma; Pathway interactions; Patients; Plasmids; Process; Production; Prognosis; Proliferating; Prostate; Proteins; Recombinants; Recurrent disease; Regenerative capacity; 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; cancer therapy; candidate identification; clinical translation; clinically relevant; comparison control; conventional therapy; cost; cytokine; depolymerization; design and construction; experience; extracellular; gemcitabine; heparanase; heparinase III; in vivo; ineffective therapies; inhibitor; macromolecule; malignant breast neoplasm; manufacture; microbial; microbial based therapy; 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; therapeutically effective; 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.

项目摘要胰腺导管腺癌（PDAC）是一种高度侵略性的疾病 预后。 PDAC的几乎宇宙致癌驱动器是小GTPase蛋白的本构激活 KRAS，诱导多个下游信号级联，共同促进快速细胞增殖， 转移和治疗性抗性。为了克服这些活动的高能量需求，克拉斯也 触发代谢适应以促进细胞外来源的营养清除，例如通过 大型细胞增多症。大型细胞增多症是一个过程，细胞外材料非特异性地吞没，并且 然后在溶酶体中降解以产生由肿瘤细胞用于生物合成的终产物。这个过程 本质上，赋予对无数合成代谢抑制剂的抗性。 syndecan-1是乙酰硫酸乙酰肝素蛋白聚糖 （HSPG）在细胞表面上调，该细胞表面是PDAC和 其他以KRAS驱动的癌症，包括膀胱，肺，前列腺，结肠和乳房。除了调解 大分子转运，HSPG可以在结合并调节的细胞外基质（ECM）中发现 许多信号分子（例如生长因子和细胞因子）与它们的同源受体的相互作用。 HSPG的亲肿瘤活性通过其乙酰肝素的酶促修饰极为调节 硫酸盐（HS）部分。哺乳动物肝素酶采用β-（1,4） - 糖苷键的水解切割 在葡萄糖酸和葡萄糖胺之间，以促进与参与的生长因子和酶的释放 ECM重塑，入侵和转移。与哺乳动物肝酶相反，细菌肝素酶III （hepiii）通过独特的β-脱离机制解散HSPG，该机制在α-（1,4） - （1,4） - 糖苷键。各种研究证实了HSPGS的HEPIII修饰抑制了新血管形成， 大型细胞增多症，肿瘤生长和转移。但是，无法将HEPIII活性限制为肿瘤组织 长期以来一直禁止其用作治疗剂。使用衰减，靶向肿瘤的沙门氏菌伤寒沙门氏菌 （st）向量，我们已经开发了第一个重组ST表达功能性HEPIII（St-Hepiii） 严格调节，可诱导的启动子。我们已经确认了St-Hepiii抑制高亲和力HS的能力 相互作用，大型细胞增多症和Kras突变肿瘤的生长。在此应用程序中，我们将：1）确定 St-Hepiii治疗对体内代谢产物可利用性和代谢相关基因途径的影响； 2） 确定合成代谢抑制剂与St-Hepiii结合使用的抗肿瘤功效； 3）发展和 表征在肿瘤可诱导型启动子下表达HEPIII的重组STS，以提高临床可行性。 完成这些目标将使我们能够开发出能够抑制肿瘤的新型靶向剂 PDAC和其他KRAS驱动癌症生存至关重要的代谢过程。我们的代理商可以使用 为了抵消对靶向合作合成代谢过程的护理标准疗法的抵抗力和 对难以治疗的癌症患者的生存产生积极影响。