P-2: Targeting telomerase expansion mechanism in MM

在上一个资助期，基于端粒功能的维持对于 为了提高MM细胞的存活率，我们研究了维持MM中端粒功能的机制，并评估了 端粒酶/端粒抑制剂作为新的治疗剂。我们已经证明，端粒酶活性是 与正常浆细胞相比，MM细胞和细胞系中端粒长度增加，而端粒长度较短， 为使用端粒维持机制导向的新疗法提供关键治疗指数。 我们进一步评估了MM中端粒酶活性的调节机制。 发现重要的MM存活信号也是维持端粒酶活性的机制。我们 已经证明，诱导MM细胞增殖和存活的IL-6和IGF-1也增加 端粒酶活性;这些细胞因子通过NF κ B介导的 hTERT mRNA和蛋白表达;以及通过Akt介导的hTERT磷酸化和 activation.我们还进一步表明，热休克蛋白90与端粒酶复合并调节端粒酶活性; 相反，17-AAG对hsp 90的抑制导致端粒酶活性的抑制。我们终于有 评估了各种端粒酶抑制剂在MM中的作用，并确定了GRN 163 L的疗效， 反义脂质化寡核苷酸hTERT抑制剂，在体外以及在体内在鼠模型中， 在该更新申请中，我们正在启动GRN 163 L在复发和复发性MM中的I期研究， 复发难治性MM，我们将评估GRN 163 L在骨髓瘤中的临床和分子作用;以及 作为鉴定在临床前体外和体内模型中与端粒酶抑制协同的药剂。的 在单药临床研究中确定的应答与耐药性的分子相关性，以及 具有协同抗MM活性的组合的临床前鉴定将为以下方面提供框架： 联合临床试验。为此，将追求以下目标： 端粒酶靶向GRN 163 L治疗复发性乳腺癌患者的安全性、有效性和分子相关性 具体目标2：确定端粒酶抑制剂和新的治疗方法的合理组合， 体外介导协同MM细胞毒性的药物;和具体目标3：确定体内疗效 端粒酶抑制剂组合疗法在人MM的鼠模型中用于翻译成衍生的 临床研究。

In the previous funding period, based on the hypothesis that maintenance of telomere function is critical to MM cell survival, we investigated the mechanisms maintaining telomere function in MM and evaluated inhibitors of telomerase/telomeres as novel therapeutics. We have shown that telomerase activity is increased, while telomere length is shorter, in MM cells and cell lines compared to normal plasma cells, providing a critical therapeutic index for using telomere maintenance mechanism-directed novel therapeutics. We have further evaluated the mechanisms regulating telomerase activity in MM. Specifically, we have identified that the important MM survival signals are also mechanisms maintaining telomerase activity. We have demonstrated that IL-6 and IGF-1, which induce MM cell proliferation and survival, also increase telomerase activity; these cytokines augment telomerase activity through NFkB-mediated upregulation of both hTERT mRNA and protein expression; as well as through Akt-mediated hTERT phosphorylation and activation. We have also gone on to show that hsp90 complexes with and modulates telomerase activity; conversely, inhibition of hsp90 by 17-AAG leads to inhibition of telomerase activity. Finally, we have evaluated the effects of various inhibitors of telomerase in MM and identified efficacy of GRN163L, an antisense lipidated oligonucleotide hTERT inhibitor, both in vitro as well as in vivo in a murine model of human MM. In this renewal application, we are initiating a phase I study of GRN163L in relapsed and relapsed refractory MM, we will evaluate the clinical and molecular effects of GRN163L in myeloma; as well as identify agents that are synergistic with telomerase inhibition in preclinical in vitro and in vivo models. The molecular correlates of response versus resistance identified in the single agent clinical study, coupled with preclinical identification of combinations with synergistic anti-MM activity, will provide the framework for combination clinical trials. To this end, the following aims will be pursued: Specific Aim 1: To investigate safety, efficacy, and molecular correlates of telomerase-targeting GRN163L therapy in patients with relapsed or refractory MM; Specific Aim 2: To define rationally-based combinations of telomerase inhibitor and novel agents which mediate synergistic MM cytotoxicity in vitro; and Specific Aim 3: To define the in vivo efficacy of telomerase inhibitor combination therapies in murine models of human MM for translation to derived clinical studies.