Development of a PIKFYVE antisense oligonucleotide treatment for FTD

开发PIKFYVE抑制性FTD反义寡核苷酸治疗 项目摘要 额颞叶痴呆（FTD）是一种复杂的疾病，由许多不同的遗传病因引起。那里 没有药物可以减缓FTD的进展。尽管针对特定病因的治疗策略 突变（例如C9 ORF 72 ASO）可能证明对个体形式的FTD有效，但这些方法不能 解决绝大多数具有未知遗传病因的病例。此外，鉴于大量 不同的基因，可能有助于FTD和事实，每种遗传形式是相对罕见的，这种策略 可能难以在所有情况下实施。因此，迫切需要新的治疗策略， 挽救多种形式的FTD，尤其是遗传病因未知的FTD。 45%的FTD患者在皮质神经元中显示TDP-43的胞质聚集，而另外45%的患者在皮质神经元中显示TDP-43的胞质聚集。 tau聚集体。研究表明，这些神经元TDP-43和tau聚集体驱动神经变性。因此，在本发明中， 为了确定FTD的新治疗靶点，我们使用细胞重编程来产生诱导的皮质 来自显示TDP-43聚集体的C9 ORF 72 FTD患者以及MAPT FTD患者的神经元（iN）， 携带tau蛋白聚集体的人然后，我们进行化学筛选，以确定拯救退化的目标， PIKFYVE激酶抑制剂是最有效的化合物之一 反义寡核苷酸（阿索）介导的PIKFYVE抑制 证实阻断PIKFYVE活性可挽救FTD iN存活。 与小分子相反，反义寡核苷酸（ASO）提供了一种简单的方法来靶向的靶向寡核苷酸。 CNS，因为它们可以直接注射到脊髓中， CNS，并且不太可能引起外周毒性。因此，我们正在寻求ASO介导的抑制， PIKFYVE作为治疗各种形式ALS的方法。我们筛选了数百名人类 PIKFYVE ASO，并在体外鉴定了10个具有有效PIKFYVE敲低的先导ASO。我们有 测试了数百个人类PIKFYVE ASO，并确定了三个有希望的开发线索。的 本直接进入II期提案的目的是进一步表征PIKFYVE的疗效 抑制，和安全的主要ASO选择一个真正的发展候选人， GLP毒性研究进展。我们发现分泌性自噬作为一种治疗方法， 神经退行性变对该领域影响很大，因为激活蛋白酶体和自噬 在神经退化模型中的结果好坏参半。

Development of a PIKFYVE suppressing antisense oligonucleotide treatment for FTD Project Summary Frontotemporal dementia (FTD) is a complex disease that results from many diverse genetic etiologies. There are no drugs that slow the progression of FTD. Although therapeutic strategies that target specific causal mutations (e.g. C9ORF72 ASOs) may prove effective against individual forms of FTD, these approaches cannot address the vast majority of cases that have unknown genetic etiology. Moreover, given the large number of different genes that likely contribute to FTD and the fact that each genetic form is relatively rare, this strategy may be difficult to implement for all cases. Thus, there is a pressing need for new therapeutic strategies that rescue multiple forms of FTD, particularly those with unknown genetic etiologies. 45% of FTD patients display cytosolic aggregates of TDP-43 in cortical neurons, while another 45% harbor tau aggregates. Studies suggest that these neuronal TDP-43 and tau aggregates drive neurodegeneration. Thus, to identify new therapeutic targets for FTD, we used cellular reprogramming to generate induced cortical neurons (iNs) from C9ORF72 FTD patients, who display TDP-43 aggregates, as well as MAPT FTD patients, who harbor tau aggregates. We then performed chemical screens to identify targets that rescue the degeneration of both C9ORF72 and MAPT FTD iNs. Inhibitors of PIKFYVE kinase were among the most potent compounds on both C9ORF72 and MAPT FTD iNs. Antisense oligonucleotide (ASO)-mediated suppression of PIKFYVE confirmed that blocking PIKFYVE activity rescues FTD iN survival. In contrast to small molecules, antisense oligonucleotides (ASOs) provide a facile approach to targeting the CNS because they can be injected directly into the spinal cord, achieve sustained target engagement throughout the CNS, and are less likely to cause peripheral toxicity. Thus, we are pursuing ASO-mediated suppression of PIKFYVE as a therapeutic approach for diverse forms of ALS. We have screened hundreds of human PIKFYVE ASOs and identified ten lead ASOs with potent PIKFYVE knockdown in vitro . We have tested hundreds of human PIKFYVE ASOs and identified three promising leads for development. The objective of this Direct to Phase 2 proposal is to further characterize the efficacy of PIKFYVE suppression, and the safety of the lead ASOs to select a bona fide development candidate for advancement in GLP toxicity studies. Our discovery of secretory autophagy as a therapeutic approach in neurodegeneration is high impact for the field because activating the proteasome and autophagy has had mixed results in neurodegeneration models.