P-5: Translation of the 3p21.3 Gene FUS1 into Pathway-Targeted Molecular Therapy

P-5：将 3p21.3 基因 FUS1 转化为通路靶向分子治疗

• 批准号: 7507387
• 负责人: Jack Roth
• 金额: $ 23.19万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-01 至 2013-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7507387
• 关键词: 3p21.3; ABL1 gene; Affect; Alleles; Apoptosis; Apoptotic; Biological Markers; Cancer Patient; Cancer cell line; Cell Cycle Arrest; Cells; Chemotherapy-Oncologic Procedure; Clinic; Clinical; Clinical Trials; Combined Modality Therapy; Data; Development; Disease regression; Distant; Encapsulated; Epidermal Growth Factor Receptor; Epithelial Cells; Erlotinib; Gefitinib; Gene Proteins; Genes; Goals; Growth; Histology; Human; Imatinib; Immunohistochemistry; In Vitro; Induction of Apoptosis; Loss of Heterozygosity; Malignant neoplasm of lung; Mediating; Molecular; Molecular Abnormality; Molecular Profiling; Molecular Target; Mus; Non-Small-Cell Lung Carcinoma; Normal Cell; Oncogenic; PDAP2 Gene; PDGFRB gene; Pathway interactions; Patients; Phase; Phase I Clinical Trials; Phenotype; Plasmids; Platinum; Population; Post-Translational Protein Processing; Primary Neoplasm; Protein Overexpression; Protein Tyrosine Kinase; Proteins; Proto-Oncogene Protein c-kit; Replacement Therapy; Resistance; Site; Specimen; Staging; Systemic Therapy; Therapeutic; Translations; Treatment Protocols; Tumor Cell Line; Tumor Suppressor Genes; Tyrosine Kinase Inhibitor; Xenograft Model; Xenograft procedure; base; c-abl Proto-Oncogenes; cancer cell; cancer gene expression; cell growth; gene function; in vivo; intravenous injection; loss of function; lung small cell carcinoma; mouse model; nanoparticle; neoplastic cell; protein expression; response; restoration; small molecule; tumor; tumor xenograft; uptake

Allelic loss and loss of function of tumor suppressor genes (TSGs) are the most frequent genetic abnormalities in lung cancer. Replacement of TSG function alone is therapeutic and often leads to lung cancer cells undergoing apoptosis or cell cycle arrest in vitro. Currently it appears that such "replacement therapy" must be done with genes rather than small-molecule mimics of TSGs. We recently demonstrated that restoration of function of 3p21.3 TSG FUS1, a proapoptotic protein, with the use of systemic nanoparticle delivery successfully cured mice with large human lung cancer orthotopic xenografts. In a phase I clinical trial systemic nanoparticle therapy delivered the FUS1 TSG to distant sites in stage IV nonsmall cell lung cancer (NSCLC) patients after intravenous injection. The FUS1 gene is inactivated in primary tumors due to 3p21.3 allele haploinsufficiency and defective post-translational modification of the remaining gene product. Enforced expression of the wild-type FUS1 in 3p21.3-deficient NSCLC cells significantly suppressed tumor cell growth by induction of apoptosis, functioning as a TSG in vitro and in vivo. However, FUS1 overexpression in human bronchial epithelial cells and other normal cells does not affect their viability. We also observed that exogenous expression of wild-type FUS1 protein in NSCLC and SCLC cells deficient in FUS1 had inhibitory effects on several oncogenic protein tyrosine kinases (PTKs), including EGFR, PDGFR, c-abl, and c-kit in NSCLC and small cell lung cancer cell lines. Associated with this PTK inhibition, there was a markedly enhanced cell response to the clinically available tyrosine kinase inhibitors (TKIs) imatinib and gefitinib. Thus, combined treatment with FUS1 and TKIs led to a significant growth inhibitory effect on lung cancer cells that were resistant to TKIs given alone. We hypothesize that treatment with the FUS1 gene delivered by nanoparticles combined with TKI therapy will have additive or supra-additive growth inhibitory and pro-apoptotic effects on lung cancer cells overcoming TKI-induced or intrinsic resistance. Our long-term goal is to develop personalized, pathway-targeted treatments which are more effective and less toxic than current treatments. The specific aims in this proposed study are 1) to determine whether FUS1-induced apoptosis and growth arrest are potentiated in various lung cancer cells /n vitro and in vivo in tumor xenograft models by TKIs that are currently being used in the clinic, 2) to identify sensitivity and resistance phenotypes associated with FUS1 expression and molecular signatures associated with these phenotypes in human lung cancer cell lines and tumor specimens and validate candidate signature molecules in a larger population of lung cancer cell lines, tumor xenografts, and clinical specimens from lung cancer patients, and 3) to conduct a Phase l/ll clinical trial combining Fl/S7-nanoparticles and erlotinib in stage IV lung cancer patients who have progressed following treatment with platinum-containing regimens.

肿瘤抑制基因（TSG）的等位基因丢失和功能丧失是最常见的遗传性肿瘤。 肺癌的异常单独替代TSG功能是治疗性的，并且常常导致肺 体外经历细胞凋亡或细胞周期停滞的癌细胞。目前看来，这种“替代 “治疗”必须用基因而不是TSGs的小分子模拟物来完成。我们最近展示了 使用全身性免疫抑制剂可以恢复促凋亡蛋白3p21.3 TSG FUS 1的功能， 纳米颗粒递送成功地治愈了具有大的人肺癌原位异种移植物的小鼠。中 I期临床试验全身性纳米颗粒治疗将FUS 1 TSG递送至IV期非小细胞肺癌的远端部位， 细胞肺癌（NSCLC）患者静脉注射后。FUS 1基因在原代细胞中失活， 肿瘤由于3p21.3等位基因单倍不足和缺陷的翻译后修饰的其余 基因产物野生型FUS 1在3p21.3缺陷型NSCLC细胞中的增强表达显著地抑制了FUS 1的表达。 通过诱导细胞凋亡抑制肿瘤细胞生长，在体外和体内起TSG的作用。然而，在这方面， FUS 1在人支气管上皮细胞和其他正常细胞中的过表达并不影响它们的活力。 我们还观察到，在NSCLC和SCLC细胞中野生型FUS 1蛋白的外源性表达， 在FUS 1中，对几种致癌蛋白酪氨酸激酶（PTK）具有抑制作用，包括EGFR， NSCLC和小细胞肺癌细胞系中的PDGFR、c-abl和c-kit。与这种PTK抑制相关， 对临床上可用的酪氨酸激酶抑制剂（TKI）的细胞应答显著增强 伊马替尼和吉非替尼。因此，FUS 1和TKI的联合治疗导致显著的生长抑制， 对TKI单独给药耐药的肺癌细胞的影响。我们假设， 通过纳米颗粒递送的FUS 1基因与TKI疗法组合将具有相加或超相加的效应。 对肺癌细胞的生长抑制和促凋亡作用克服TKI诱导的或内在的 阻力我们的长期目标是开发个性化的，有针对性的治疗方法， 比目前的治疗方法更有效，毒性更低。本研究的具体目标是：1）确定 FUS 1诱导的细胞凋亡和生长停滞是否在各种肺癌细胞中增强/体外， 通过目前临床上使用的TKI在肿瘤异种移植模型中进行体内试验，2）确定敏感性 和与FUS 1表达相关的抗性表型以及与 这些表型在人肺癌细胞系和肿瘤标本中的表达，并验证候选签名 肺癌细胞系、肿瘤异种移植物和来自肺的临床标本的较大群体中的分子 癌症患者，和3）进行组合F1/S7-纳米颗粒和厄洛替尼的I/II期临床试验， 在用含铂方案治疗后进展的IV期肺癌患者。