Cyclin Kinase Inhibitors as Molecular Targets for Cancer

描述（由申请人提供）： 这是一个重新提交的申请，其重点是细胞周期蛋白激酶 CKI抑制剂作为癌症治疗的新的潜在靶点。而 直到最近，这种模式一直认为这些蛋白质只作为 细胞生长抑制剂，我们最近发表的工作表明，CKI p21 Wafl/Cipl在某些条件下和某些细胞类型下， 对血管平滑肌（VSM）细胞生长的影响，使我们提出 这类分子是癌症化疗中的新的潜在靶点。 自首次提交这一申请以来，我们获得了新的数据 进一步验证该分子作为这样的靶标。此外，我们还有更多 新的（而且，我们谦虚地认为，非常令人兴奋的）数据提供了证据， 用我们简单反义寡脱氧核苷酸（oligo）抑制p21 转染技术导致肿瘤血管生成的显著抑制， 以及肿瘤生长。的 我们希望在这项研究中产生的兴奋，部分原因是 易于给药，没有毒性，没有表型 p21敲除的变化（表明p21作为一种免疫抑制剂的信号传导中的冗余）。 肿瘤抑制因子p53的效应子），以及整个动物的可能性。 这种新的反义寡核苷酸技术针对新靶点的应用 分子用于治疗一种毁灭性的人类疾病。 根据收到的批评意见，对重新提交的文件进行了修改 从上次提交，通过删除许多机械的 实验和缩小工作的重点，以一个单一的CKI，p21， 我们掌握的数据最多。更重要的是， 在这个重新提交支持我们的假设，我们现在建议， 仔细研究一下我们发现了一种令人兴奋的可能性， 在肿瘤中作用于VSM细胞支架的抗血管生成分子 血管生成血管具体目标是（1）确定最佳的 反义CKI寡核苷酸是生长抑制性、促增殖性或 抗血管生成;（2）确定p21反义寡核苷酸局部作用 对肿瘤细胞生长的影响，并在全身注射转移性 开始研究抗血管生成的机制 反义p21寡核苷酸的作用。 我们相信，完成这一应用的实验将充分 验证p21反义寡核苷酸在哺乳动物癌症领域中的用途 并且不仅将导致提交可资助的R 01申请的足够数据， 但更重要的是，进一步的动物试验， 抑制作为人类癌症中可行的治疗辅助物。

DESCRIPTION (provided by applicant): This is a resubmission of a application which focuses on the cyclin kinase inhibitors (CKIs) as novel potential targets for cancer therapy. While the paradigm, until quite recently, has been that these proteins act solely as inhibitors of cell growth, our recent published work showing that the CKI p21 Wafl/Cipl has, under some conditions and with some cell types, permissive effects on vascular smooth muscle (VSM) cell growth, has led us to propose this class of molecules as novel potential targets in cancer chemotherapy. Since the first submission of this application, we have obtained new data further validating this molecule as such a target. Furthermore, we have more new (and, we modestly believe, very exciting) data providing evidence that inhibition of p21 by our simple antisense oligodeoxynucleotide (oligo) transfection technique leads to marked inhibition of tumor angiogenesis, as well as tumor growth, in an in vivo animal model of mammary cancer. The excitement which we hope to generate in this research is, in part, due to the ease of administration, the lack of toxicity, the absence of phenotypic changes in p21 knockouts (suggesting redundancy in signaling of p21 as an effector of the tumor suppressor p53), and the possibility for whole animal application of this novel antisense oligo technique against a novel target molecule for treatment of a devastating human disease. This resubmission has been modified, in response to the critiques received from the last submission, by the removal of many of the mechanistic experiments and a narrowing of the focus of the work to a single CKI, p21, concerning which we have the most data. Significantly more preliminary data is provided in this resubmission supporting our hypothesis, and we now propose to examine in some detail the exciting possibility that we have discovered an antiangiogenic molecule that is acting upon the VSM cell scaffolding in tumor angiogenic vessels. The Specific Aims are (1) to determine optimal by which the antisense CKI oligos are growth inhibitory, apoptosis-promoting, or anti-angiogenic; (2) to determine the effect of p21 antisense oligos locally on tumor cell growth and systemically in a mouse injected with metastatic tumor cells; and (3) to begin to study the mechanism of the anti-angiogenic effect of antisense p21 oligos. We believe that completion of the experiments in this application will fully validate the use of antisense oligos to p21 in the field of mammalian cancer and will lead not only to adequate data to submit a fundable R01 application, but, more importantly, to further animal, and ultimately human, trials of p21 inhibition as a viable treatment adjunct in human cancer.