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 疼痛性椎间盘（IVD）退变（IDD）的快速增加使其迫切需要开发 延缓IDD进展的解决方案。IDD与慢性炎症、IVD细胞结构丧失、 基质降解和细胞凋亡，这是由AKT，PKC和MAPK的去磷酸化加速的 信号通路我们提出的初步证据表明，磷酸酶Pleckstrin同源结构域 富亮氨酸重复蛋白磷酸酶1（PHLPP 1）促进IDD。因此，PHLPP 1的整体缺失 通过防止基质降解和减少促炎性因子抑制老年小鼠IDD进展 细胞因子表达我们对退化的人类NP细胞的初步数据表明，促炎性细胞因子 在用小分子PHLPP抑制剂治疗后，反应降低。我们之前开发了 可注射的纳米制剂，其能够在长时间内以恒定速率递送小分子， 段时间基于这些发现，我们的初步概念和可行性证明，以及我们最近的 出版物，我们建议测试新的假设，即NP隔室特异性抑制PHLPP 1， 通过抑制炎症和基质降解延缓疾病进展 IDD。 目的1将使用年龄诱导的IDD模型，在体内确定PHLPP 1在NP隔室中对IDD的作用。 条件Phlpp1基因敲除小鼠自发性IDD。Phlpp1将在NP（NPcKO）中被耗尽， Phlpp1的免疫组化，分子和生物力学方法进行评估。体外研究 将通过单细胞RNA测序和分子生物学方法评估差异调节途径。 小鼠NPcKO以及人NP细胞中PHLPP 1敲低的分析。目标2将测试 小分子PHLPP抑制剂通过开发可长期注射纳米制剂来减缓IDD PHLPP抑制剂释放和评估其在自发IDD小鼠模型中的功效。 这个项目是非常重要的，因为椎间盘源性背痛是美国的一个主要负担。新 IDD的机制和微创治疗的见解将联合收割机的机制和翻译结合起来， 问题研究这项研究是创新性的，因为小分子PHLPP抑制剂的开发还没有完成。 以及NIR标记的纳米制剂用于控制PHLPP抑制剂递送至 治疗IDD是一项技术创新。本研究的成功完成将为NP提供新的见解 IDD隔室特异性进展和高级小分子PHLPP抑制剂负载纳米制剂 注射作为人类IDD的潜在疾病缓解治疗。