Targeting PHLPP to treat interval disc degeneration using surgical and drug delivery methods

使用手术和药物递送方法靶向 PHLPP 治疗间盘退变

• 批准号: 10367568
• 负责人: Svenja Illien-Junger
• 金额: $ 40.88万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-10 至 2027-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10367568
• 关键词: Address; Aftercare; Age; Aging; Animal Model; Apoptosis; Back Pain; Biochemical; Biological; Biomechanics; Cell Death; Cell Proliferation; Cells; Cellularity; Chronic; Clinical; Data; Degenerative polyarthritis; Deposition; Deterioration; Development; Disease; Disease Progression; Drug Delivery Systems; Ensure; Enzyme-Linked Immunosorbent Assay; Enzymes; Extracellular Matrix; Extracellular Matrix Degradation; Formulation; Genes; Genotype; Glycosaminoglycans; Human; In Vitro; Inflammation; Inflammatory; Inflammatory Response; Injectable; Injections; Intervertebral disc structure; Knock-out; Knockout Mice; Label; Lead; MAP Kinase Gene; MAPK Signaling Pathway Pathway; Methods; Modality; Modeling; Molecular; Mus; Musculoskeletal; Nanotubes; Needles; Operative Surgical Procedures; Osteoclasts; Outcome Measure; PH Domain; PI3K/AKT; Pain; Pathway interactions; Pattern; Pharmaceutical Preparations; Phosphoric Monoester Hydrolases; Phosphorylation; Play; Property; Protein Dephosphorylation; Protein phosphatase; Proto-Oncogene Proteins c-akt; Public Health; Publications; Publishing; Research; Role; Signal Pathway; Signal Transduction; Small Interfering RNA; Suspensions; Testing; Therapeutic; Time; Tissues; Transgenic Organisms; United States; Western Blotting; Wild Type Mouse; Work; aged; base; biomaterial compatibility; bone; cytokine; delivery vehicle; effective therapy; efficacy evaluation; efficacy testing; experimental study; fetal; human tissue; improved; in vivo; inflammatory marker; inhibitor; innovation; insight; interest; intervertebral disk degeneration; knock-down; leucine-rich repeat protein; loss of function; macrophage; minimally invasive; mouse model; mutant; nanocarrier; nanoformulation; nanoparticle; neutrophil; novel; nucleus pulposus; pain relief; prevent; single-cell RNA sequencing; small molecule; small molecule inhibitor; socioeconomics; transcriptome sequencing; translational study

PROJECT SUMMARY The rapid increase in painful intervertebral disc (IVD) degeneration (IDD) makes it an urgent need to develop solutions for delaying IDD progression. IDD is associated with chronic inflammation, loss of IVD cellularity, matrix degradation and apoptosis, which are accelerated by dephosphorylation of AKT, PKC, and MAPK signaling pathways. We present preliminary evidence that the phosphatase Pleckstrin homology domain leucine-rich repeat protein phosphatase 1 (PHLPP1) promotes IDD. Thus global deletion of PHLPP1 repressed IDD progression in old mice by preventing matrix degradation and reducing pro-inflammatory cytokine expression. Our preliminary data on degenerated human NP cells suggested that pro-inflammatory responses were decreased after treatment with a small molecule PHLPP inhibitor. We previously developed injectable, nanoformulations that are capable of delivering small molecules at constant rates over an extended period of time. Based on these findings, our preliminary proof of concept and feasibility as well as our recent publication, we propose to test the novel hypothesis that NP compartment specific inhibition of PHLPP1 will delay disease progression via suppressing inflammation and matrix degradation in age-induced spontaneous IDD. Aim 1 will identify the role of PHLPP1 on IDD in the NP compartment in vivo using a model of age-induced spontaneous IDD in conditional Phlpp1 knockout mice. Phlpp1 will be depleted in NP (NPcKO) and the role of Phlpp1 will be assessed with immunohistochemical, molecular, and biomechanical methods. In vitro studies will evaluate the differentially regulated pathways by single cell RNA-sequencing and molecular-biological analysis of mouse NPcKO as well as PHLPP1 knockdown in human NP cells. Aim 2 will test the efficacy of a small molecule PHLPP inhibitor to decelerate IDD by developing an injectable nanoformulation for long-term PHLPP inhibitor release and evaluate its efficacy in a mouse model of spontaneous IDD. This project is highly significant because discogenic backpain is a major burden in the United States. New insights in mechanisms and minimal invasive treatments of IDD will combine mechanistic and translational studies. This study is innovative because the development of a small molecule PHLPP inhibitor has not been previously described and the use of NIR labeled nanoformulations for controlled PHLPP inhibitor delivery to treat IDD is a technical innovation. Successful completion of this study will provide new insights into NP compartment specific progression of IDD and advance small molecule PHLPP inhibitor laden nanoformulation injection as a potential disease-modifying treatment for IDD in humans.

项目摘要 痛苦的椎间盘（IVD）变性（IDD）的迅速增加使其迫切需要发展 延迟IDD进程的解决方案。 IDD与慢性炎症，IVD细胞丧失有关， 基质降解和凋亡，通过Akt，PKC和MAPK的去磷酸化加速 信号通路。我们提供了磷酸酶Pleckstrin同源域的初步证据 富含亮氨酸的重复蛋白磷酸酶1（PHLPP1）促进IDD。因此，全球删除PHLPP1 通过防止基质降解和减少促炎性的抑制旧小鼠的IDD进展 细胞因子表达。我们关于退化的人NP细胞的初步数据表明促炎性 用小分子PHLPP抑制剂处理后，反应降低。我们以前开发了 可注射的，能够以延长的速度传递小分子的纳米成型 一段时间。基于这些发现，我们的初步概念证明和可行性以及我们最近的 出版物，我们建议检验新的假设，即NP隔室特定于PHLPP1的抑制作用将 通过抑制年龄引起的自发性炎症和基质降解，延迟疾病进展 IDD。 AIM 1将使用年龄诱导的模型来确定PHLPP1在体内NP隔室中IDD的作用 有条件的PHLPP1敲除小鼠中的自发IDD。 PHLPP1将在NP（NPCKO）中耗尽，并且角色 PHLPP1的of将通过免疫组织化学，分子和生物力学方法进行评估。体外研究 将通过单细胞RNA测序和分子生物学评估差异调节的途径 小鼠NPCKO以及人NP细胞中PHLPP1敲低的分析。 AIM 2将测试一个 小分子pHLPP抑制剂通过开发可长期注射的纳米制剂来减速IDD PHLPP抑制剂释放并评估其在自发IDD的小鼠模型中的功效。 该项目非常重要，因为盘源背端是美国的重大负担。新的 IDD的机制和最小侵入性治疗的见解将结合机械和翻译 研究。这项研究具有创新性，因为小分子PHLPP抑制剂的发展尚未 先前描述的以及使用NIR标记的纳米成型用于受控的PHLPP抑制剂递送到 Treat IDD是一项技术创新。这项研究的成功完成将为NP提供新的见解 室室的特异性进展和前进的小分子pHLPP抑制剂laden纳米取样 注射作为人类IDD的潜在疾病改良治疗。