A novel bone-targeting AAV-mediated gene therapy to promote bone formation in osteoporosis

一种新型骨靶向 AAV 介导的基因疗法可促进骨质疏松症的骨形成

• 批准号: 10458098
• 负责人: Jae-Hyuck Shim
• 金额: $ 21.89万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10458098
• 关键词: Adaptor Signaling Protein; Adult; Adverse effects; Affinity Chromatography; Age; Aging; American; Anabolic Agents; Angiogenic Factor; Antibodies; Binding Sites; Biodistribution; Biological; Bone Density; Bone Resorption; Bone necrosis; Bone structure; Candidate Disease Gene; Capsid; Cell Lineage; Cells; Clinical; Data; Deterioration; Disease; Endothelium; Engineering; Equilibrium; Estrogens; FDA approved; Femoral Fractures; Gene Transduction Agent; Gene Transfer; Health; Histology; Histopathology; In Vitro; Infection; Injections; Intravenous; Jaw; Label; Liver; Mass Spectrum Analysis; Measures; Mediating; MicroRNAs; Modification; Mus; Myocardium; Operative Surgical Procedures; Osteoblasts; Osteoclasts; Osteocytes; Osteogenesis; Osteoporosis; Osteoporotic; Ovariectomy; PTH gene; Pathogenicity; Pathway interactions; Patients; Phenotype; Postmenopausal Osteoporosis; Proteins; Proteomics; RNA Interference; Recombinant adeno-associated virus (rAAV); Repression; Reverse Transcriptase Polymerase Chain Reaction; Senile Osteoporosis; Serotyping; Serum Calcium Level; Skeletal Muscle; Therapeutic; Therapeutic Agents; Therapeutic Intervention; Tissues; Tropism; Validation; Viral Vector; WNT Signaling Pathway; aged; antagonist; base; biomechanical test; bone; bone aging; bone loss; bone mass; calcification; fracture risk; gene therapy; immunogenicity; improved; in vivo; innovation; knock-down; long bone; mechanical properties; mouse model; new therapeutic target; novel; optical imaging; osteogenic; osteoporosis with pathological fracture; parathyroid hormone-related protein; response; side effect; skeletal; spine bone structure; targeted treatment; therapeutic gene; therapeutic target; transcriptome; transduction efficiency; transgene expression; vector

PROJECT SUMMARY Adult bone mass is determined by the balance between bone formation by osteoblasts (OBs) and bone resorption by osteoclasts (OCs), and disturbances in this equilibrium to favor osteoclast (OC)-mediated resorption leads to osteoporosis. The majority of existing therapeutics for osteoporosis act by inhibiting OCs, but these can not cure osteoporosis and are limited by rare side effects. Current anabolic agents, parathyroid hormone (PTH), parathyroid hormone-related protein (PTHrP), and anti-sclerostin antibody exist for promoting osteoblast (OB) function to treat patients with osteoporosis. However, these agents are also limited by concern for off-target adverse effects and waning efficacy. Previously, we and others demonstrated that inhibition of potent OB suppressors, sclerostin (SOST) and the adaptor protein schnurri-3 (SHN3), promotes bone formation in mouse models of postmenopausal and senile osteoporosis. One innovative approach to treat osteoporosis is RNAi-based bone anabolic gene therapy using recombinant adeno-associated virus (rAAV). Using our engineered rAAV9 vector with bone-specific tropism and transgene expression, we will develop novel gene therapeutics that promote bone formation in osteoporosis with a single systemic administration. Additionally, we will identify potential novel osteogenic and/or angiogenic factors regulated by the SOST/SHN3 pathway that could be useful as therapeutic targets for osteoporosis. Aim 1 will examine whether bone-specific rAAV9-mediated silencing of SHN3 or SOST can reverse bone loss in mouse models of osteoporosis. To avoid potential off-target adverse effects in non-skeletal tissues, rAAV9’s bone- specific tropism and transgene expression were further improved by capsid modification and tissue-specific miRNA-mediated repression of rAAV expression. Using two mouse models of osteoporosis (ovariectomized (OVX) and aged mice), we will determine therapeutic potentials of rAAV9-mediated silencing of SHN3 or SOST in postmenopausal and senile osteoporosis. Aim 2 will identify novel osteogenic and/or angiogenic factors regulated by the SHN3/SOST pathway that promote bone formation in osteoporosis. In vivo silencing accuracy of rAAV9 carrying amiR-shn3 or amiR-sost in bone-residing OB lineage cells will be examined by scatterplot analysis of whole transcriptome data. Once validated, wt, shn3- or sost-deficient OB-lineage cells will be FACS-sorted from AAV-treated, OVX-mice and subjected for transcriptome profiling. As a complimentary approach, proteomics will be performed in AAV-transduced OB-lineage cells isolated from MetRS;Prx1-cre mice with OB-specific labeling of nascent proteins after OVX-surgery. These combinatory approaches will allow us to identify novel osteogenic and/or angiogenic factors commonly or differentially regulated by the pathways of SHN3 and SOST in response to estrogen deficiency-induced osteoporosis. Successful completion of these aims will provide proof-of-concept demonstration and identify potential novel regulators that could be useful as therapeutic targets for osteoporosis.

项目摘要 成人骨骼质量取决于成骨细胞（obs）和骨之间的骨骼形成之间的平衡 破骨细胞（OCS）的分辨率，以及在这种等同的灾难中，以偏爱破骨细胞（OC）介导的 分辨率导致骨质疏松症。大多数现有的骨质疏松症疗法通过抑制OC， 但是，这些无法治愈骨质疏松症，并且受到罕见副作用的限制。当前的合成代理，甲状旁腺 马酮（PTH），甲状旁腺与马匹相关蛋白（PTHRP）和抗骨蛋白抗体存在用于促进 成骨细胞（OB）功能以治疗骨质疏松症患者。但是，这些代理也受到关注的限制 为了脱离目标的不良反应和减弱的有效性。以前，我们和其他人证明了对 有效的OB补充剂，硬化蛋白（SOST）和衔接蛋白Schnurri-3（SHN3）促进骨骼 绝经后和老年骨质疏松症的小鼠模型中的形成。一种创新的治疗方法 骨质疏松症是基于RNAi的骨合成基因疗法，使用重组腺相关病毒 （拉夫）。使用我们工程的RAAV9载体与骨特异性的偏移和转化表达，我们将 开发新型基因疗法，促进具有单一系统性的骨质疏松症的骨形成 行政。此外，我们将确定由受调节的潜在新型成骨和/或血管生成因子 可以用作骨质疏松症的治疗靶标的SOST/SHN3途径。 AIM 1将检查 骨特异性RAAV9介导的SHN3的沉默还是SOST可以逆转小鼠的骨质流失 骨质疏松症的模型。为了避免在非骨骼组织中潜在的脱靶不良反应，Raav9的骨 特异性的向向向向潮流和转化表达通过衣壳修饰和组织特异性进一步改善 miRNA介导的RAAV表达的表示。使用两种骨质疏松症的小鼠模型（Ovariectomized （OVX）和老年小鼠），我们将确定RAAV9介导的SHN3或SOST的静音的治疗潜力 在绝经后和老年骨质疏松症中。 AIM 2将识别新颖的成骨和/或血管生成因子 受SHN3/SOST途径的调节，该途径促进骨质疏松症的骨形成。体内沉默 Raav9携带Amir-SHN3或在骨固定OB谱系中携带Amir-SHN3或Amir-Sost的准确性将通过 整个转录组数据的散点图分析。一旦验证，WT，SHN3-或SOST缺陷的OB-LINEGE细胞 将从经AAV处理的OVX鼠标中进行FACS分类，并进行转录组分析。作为 免费方法，将在从中分离出的AAV转导的OB线细胞中进行蛋白质组学 OVX手术后，具有新生蛋白的ob特异性标记的metrs; prx1-cre小鼠。这些组合 方法将使我们能够识别出通常或不同的新型成骨和/或血管生成因子 响应于雌激素缺乏引起的骨质疏松症的响应，受SHN3和SOST的途径调节。 这些目标的成功完成将提供概念验证示范并确定 潜在的新型调节剂可以用作骨质疏松症的治疗靶标。

项目成果

Biphasic regulation of osteoblast development via the ERK MAPK-mTOR pathway.

• DOI: 10.7554/elife.78069
• 发表时间: 2022-08-17
• 期刊: ELIFE
• 影响因子: 7.7
• 作者: Kim, Jung-Min;Yang, Yeon-Suk;Hong, Jaehyoung;Chaugule, Sachin;Chun, Hyonho;van Der Meulen, Marjolein C. H.;Xu, Ren;Greenblatt, Matthew B.;Shim, Jae-Hyuck
• 通讯作者: Shim, Jae-Hyuck

AAV-mediated delivery of osteoblast/osteoclast-regulating miRNAs for osteoporosis therapy.

• DOI: 10.1016/j.omtn.2022.07.008
• 发表时间: 2022-09-13
• 期刊: MOLECULAR THERAPY NUCLEIC ACIDS
• 作者: John, Aijaz Ahmad;Xie, Jun;Yang, Yeon-Suk;Kim, Jung-Min;Lin, Chujiao;Ma, Hong;Gao, Guangping;Shim, Jae-Hyuck
• 通讯作者: Shim, Jae-Hyuck

Impaired mitochondrial oxidative metabolism in skeletal progenitor cells leads to musculoskeletal disintegration.

• DOI: 10.1038/s41467-022-34694-8
• 发表时间: 2022-11-11
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Lin, Chujiao;Yang, Qiyuan;Guo, Dongsheng;Xie, Jun;Yang, Yeon-Suk;Chaugule, Sachin;DeSouza, Ngoc;Oh, Won-Taek;Li, Rui;Chen, Zhihao;John, Aijaz A.;Qiu, Qiang;Zhu, Lihua Julie;Greenblatt, Matthew B.;Ghosh, Sankar;Li, Shaoguang;Gao, Guangping;Haynes, Cole;Emerson, Charles P.;Shim, Jae-Hyuck
• 通讯作者: Shim, Jae-Hyuck

Gene Therapy for Fibrodysplasia Ossificans Progressiva: Feasibility and Obstacles.

• DOI: 10.1089/hum.2022.023
• 发表时间: 2022-08
• 期刊: HUMAN GENE THERAPY
• 影响因子: 4.2
• 作者: Eekhoff, Elisabeth M. W.;de Ruiter, Ruben D.;Smilde, Bernard J.;Schoenmaker, Ton;de Vries, Teun J.;Netelenbos, Coen;Hsiao, Edward C.;Scott, Christiaan;Haga, Nobuhiko;Grunwald, Zvi;De Cunto, Carmen L.;di Rocco, Maja;Delai, Patricia L. R.;Diecidue, Robert J.;Madhuri, Vrisha;Cho, Tae-Joon;Morhart, Rolf;Friedman, Clive S.;Zasloff, Michael;Pals, Gerard;Shim, Jae-Hyuck;Gao, Guangping;Kaplan, Frederick;Pignolo, Robert J.;Micha, Dimitra
• 通讯作者: Micha, Dimitra