APOPTOSIS INDUCING GENE THERAPY FOR ORAL PREMALIGNANCY

口腔鳞状细胞癌的患者 患有恶性肿瘤，经常出现晚期疾病， 管理往往对这些人的言论产生深远的影响， 吞咽，外观，以及生存。 不幸的是， 尽管当代外科手术和放射治疗在过去的10年中取得了进步， 在过去的30年里，这些患者的生存率基本上保持不变， 没有改变 此外，化疗在癌症中几乎没有表现出希望。 管理这些癌症。 因此，开发新的机制， 阐明导致这些恶性肿瘤的癌前过程可能 在这一医学领域取得了最重大的进步。 口腔癌前病变和原位癌的基因治疗 可能是非常可行的。 上皮的局部转导， 在癌前病变过程中发生的转化很可能是 灵验。 对野生型p53腺病毒的初步研究表明， 由于其作为肿瘤的功能， 抑制基因和细胞凋亡的作用。 然而，重要的 需要在理解转导的优化方面取得进展， 分子治疗的效率，以及增加我们对 分子干预策略的相互作用， 肿瘤细胞死亡和非恶性肿瘤保留的预期效果 细胞 我们选择使用腺病毒载体作为一种机制， 由于其特征性瞬时表达的分子干预， 表达效率和扩增至高滴度的能力。 此外，从生物安全性角度来看，缺乏永久性整合是有利的。 与逆转录病毒载体相比。 因此，这项建议 为进一步加深我们对凋亡诱导基因的理解， 癌前病变和微浸润性鳞状细胞癌的治疗 口腔通过以下具体目标： 1. 确定腺病毒载体的最大转染效率 在器官型和组织中标记基因的瞬时表达中 口腔癌前病变的切片模型。 2. 为了确定野生型瞬时表达的有效性， p53、p16、p21（CIP/WAF 1）和pRB 105独立地和组合地 诱导口腔鳞癌细胞系凋亡的研究 癌前病变模型。 3. 为了确定转录因子的调节，包括 p53下游的p21（CIP/WAF 1）、E2 F-1和DP-1可进一步 增强口腔鳞状细胞癌细胞凋亡的诱导， 癌前病变的模型。 4. 为了确定优化的基因治疗策略对 诱导癌前口腔癌细胞凋亡， 正常口腔上皮和基质细胞。

Patients with squamous cell carcinoma of the oral cavity are afflicted with a malignancy that frequently presents with advanced disease and whose management often has profound effects upon these individual's speech, swallowing, cosmetic appearance, as well as survival. Unfortunately, despite advances in contemporary surgery and radiation therapy over the past thirty years, survival among these patients has remained essentially unaltered. Additionally, chemotherapy has shown little promise in the management of these cancers. Therefore, developing novel mechanisms of cessating the premalignant processes which lead to these malignancies may provide the most significant advancements in this field of medicine. Gene therapy for premalignancy and carcinoma in-situ of the oral cavity may be very achievable. Topical transduction of epithelium which has undergone transformation in the premalignant process is likely to be efficacious. Initial investigations of wild-type p53 adenovirus has shown promise as a molecular intervention due to its function as a tumor suppressor gene and proposed role in apoptosis. However, significant advances are required in understanding the optimization of transduction efficiency in molecular therapy as well as increasing our understanding of the interaction of molecular intervention strategies in order to produce the desired affect of tumor cell demise and sparing of non-malignant cells. We have elected to employ the adenovirus vector as a mechanism for molecular intervention due to its characteristic transient expression, efficiency of expression, and ability to expand to high titers. Additionally, lack of permanent integration is favorable from a biosafety standpoint as compared to retrovirus vectors. This proposal therefore submits to further develop our understanding of apoptosis inducing gene therapy for premalignant and microinvasive squamous cell carcinoma of the oral cavity through the following specific aims: 1. To determine the maximal transfection efficiency of adenovirus vectors in the transient expression of marker genes in organotypic and tissue slice models of premalignancies of the oral cavity. 2. To determine the effectiveness of transient expression of wild-type p53, p16, p21 (CIP/WAF1), and pRB105 independently and in combination in the induction of apoptosis in oral squamous carcinoma cell lines and the models of premalignancy. 3. To determine whether regulation of transcription factors, including p21 (CIP/WAF1), E2F-1, and DP-1, which are downstream of p53, can further potentiate the induction of apoptosis in oral squamous carcinoma cells and the models of premalignancy. 4. To determine the effect of the optimized gene therapy strategies for induction of apoptosis, in premalignant oral cavity carcinomas, upon normal oral epithelium and stromal cells.