A Novel Agent to Suppress Tumor Growth in Bone, Prevent Cachectic Muscle Loss and Preserve Skeletal Integrity

一种抑制骨肿瘤生长、防止恶病质肌肉损失并保持骨骼完整性的新型药物

• 批准号: 9912632
• 负责人: JOHN M CHIRGWIN
• 金额: --
• 依托单位: RLR VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9912632
• 关键词: Adipocytes; Age; Agonist; Automobile Driving; Biological Assay; Biology; Bone Diseases; Bone Pain; Bone Regeneration; Bone Resorption; Bone remodeling; Breast; Breast Cancer Cell; Breast Cancer Model; Cachexia; Cancer Control; Cancer Etiology; Cartoons; Cattle; Cell Line; Cells; Clinical; Clinical Trials; Coculture Techniques; Coupled; Cyclic AMP; Data; Disseminated Malignant Neoplasm; Fatty acid glycerol esters; Financial Hardship; GTP-Binding Proteins; Genes; Growth; Health; Human; Hypercalcemia; Hypercalcemia of Malignancy; Immunocompetent; In Vitro; Incidence; Individual; Lead; Ligands; Luciferases; Lytic Metastatic Lesion; Malignant Bone Neoplasm; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of prostate; Metastatic Neoplasm to the Bone; Modeling; Multiple Myeloma; Mus; Muscle; Muscular Atrophy; Myalgia; Neoplasm Metastasis; Organ; Osteoblasts; Osteoclasts; Osteocytes; Osteogenesis; Osteolysis; Osteolytic; Osteoporosis; Pain; Parathyroid Neoplasms; Pathological fracture; Patients; Peptides; Preclinical Testing; Prevention; Production; Prostate; Quality of life; Sensory; Signal Transduction; Skeletal Muscle; Societies; TNFSF11 gene; Testing; Transforming Growth Factor beta; Tumor Burden; Tumor-Secreted Protein; Xenograft Model; Yin; advanced breast cancer; analog; angiogenesis; beta-arrestin; bone; bone cell; bone health; bone loss; bone preservation; cancer cachexia; cancer cell; cancer clinical trial; cancer site; clinically relevant; improved; in vivo; malignant breast neoplasm; mouse model; neoplastic cell; nerve supply; neutralizing antibody; novel; osteosarcoma; parathyroid hormone-related protein; patient population; prevent; receptor; response; skeletal; skeletal muscle wasting; skeletal preservation; success; targeted treatment; tumor; tumor growth

ABSTRACT: Over 85% of patients with advanced cancers of breast and prostate suffer from incurable bone metastases. Osteoclast activation and osteoblast suppression lead to bone loss, pathologic fractures, hypercalcemia and bone pain, often accompanied by loss of skeletal muscle (cachexia). Current therapies target osteoclasts and have limited effects on tumor growth, while not restoring lost bone or muscle. Tumor-secreted parathyroid hormone-related protein, PTHrP, is a major causal agent of bone destruction by inducing expression of RANKL to increase osteoclast number and function. TGFβ, released during bone resorption, stimulates breast cancer production of PTHrP [Yin et al, 1999], driving a vicious cycle of bone metastases [Chirgwin & Guise, 2000; Kakonen et al, 2002]. PTHrP stimulates growth of PTH1R receptor- expressing tumors and indirectly causes cancer cachexia via reprogramming of adipocytes [Kir & Spiegelman, 2016]. It causes hypercalcemia of malignancy, stimulates angiogenesis and cancer bone pain. We believe the deleterious actions of tumor PTHrP can be blocked by a unique PTH1R ligand, while having beneficial anabolic effects on bone - which cannot be achieved with PTHrP-neutralizing antibody. [D-Trp12,Tyr34]-bovine-PTH(7- 34), bPTH7-34DD, is a unique inverse agonist of G protein-coupled signaling by PTH1R, decreasing cAMP, while activating β-arrestin signaling. It increases bone formation without causing hypercalcemia or osteolysis [Maudsley et al, 2015]. Our data showed anti-osteolytic and anti-tumor actions of bPTH7-34DD in co-cultures of breast cancer with mouse bone. It also countered adipocyte gene changes associated with cancer cachexia. We hypothesize that bPTH7-34DD will: 1) Inhibit the vicious cycle of osteolytic bone metastases by suppressing osteolysis & tumor growth; 2) Decrease cancer cachexia by blocking white-to-brown adipocyte conversion in bone; 3) Preserve bone health by stimulating new bone formation; 4) Effectively treat osteolytic tumors due breast cancer. 5) Be effective against other PTHrP-secreted tumor types. bPTH7-34DD will be tested in xenograft models of breast cancer bone metastasis, in prevention and treatment settings. The mechanism of bPTH7-34DD signaling will be tested in ex vivo co-cultures of cancer cells with mouse bones and in bone and fat cell lines and 1o cells and with bones and cells from wt & β-arrestin k/o (βarr2-/-) mice. Specific Aim 1: a) Determine growth effects ex vivo of bPTH7-34DD on tumor cells; b) Determine the mechanism of action of on i) Osteoblasts (Obs), osteoclasts (Ocs), & osteocytes (Ots); ii) On adipocytes; c) Test if responses to bPTH7-34DD require β-arrestin, using bones & cells from βarr2-/- mice; d) Test if bPTH7-34DD blocks growth in bone and osteolysis ex vivo with prostate and lung cancer and multiple myeloma cells Specific Aim 2: Test bPTH7-34DD in vivo for significant effects on: a) Tumor burden; b) Osteolytic lesions; c) Bone loss/formation systemically and adjacent to tumor; d) Skeletal muscle. Significance: bPTH7-34DD could be rapidly brought into clinical trials for cancer bone metastasis. Similar PTH & PTHrP peptides are used to treat osteoporosis. Benefit may extend to other PTHrP-expressing cancers that metastasize to bone, including multiple myeloma, which also colonizes bone and causes osteolysis. Technical Approaches: The project uses co-cultures of cancer cells with mouse bones, called EVOCA (ex vivo organ co-culture assay). Tumor growth is assayed by secreted Gaussia luciferase (gLuc) from stably transduced cancer cells. Changes in bone and MM genes are quantified by mouse- and human-specific qPCR. Two mouse models of breast cancer bone metastases will be used for preclinical testing. Consultants: Dr. Louis Luttrell is a world expert in PTH signaling and deciphered the actions of bPTH7-34DD. Dr. Teresa Zimmers is an expert in cancer cachexia. Dr. Jesus Delgado-Calle is an expert in osteocyte biology and osteocyte:adipocyte interactions in myeloma. Dr. David Roodman is a world expert on myeloma bone disease.

摘要：超过85％的乳房和前列腺晚期癌症患者患有无法治愈的骨骼 转移。破骨细胞活化和成骨细胞抑制导致骨质流失，病理骨折， 高钙血症和骨痛，通常伴随着骨骼肌的丧失（恶病质）。当前的疗法靶标 破骨细胞的影响有限，对肿瘤生长的影响有限，同时又不恢复骨骼或肌肉损失。 肿瘤分泌的甲状旁腺激素相关蛋白PTHRP是骨破坏骨的主要因果。 诱导RANKL的表达增加破骨细胞的数量和功能。 TGFβ，在骨骼中释放 分解，刺激PTHRP的乳腺癌产生[Yin等，1999]，驱动骨骼的恶性循环 转移[Chirgwin＆Guise，2000年； Kakonen等，2002]。 PTHRP刺激PTH1R受体的生长 通过重新编程脂肪细胞[Kir＆Spiegelman， 2016]。它会引起恶性肿瘤高钙血症，刺激血管生成和癌症骨痛。我们相信 肿瘤PTHRP的有害作用可以被独特的PTH1R配体阻止，同时具有有益的合成代谢 对骨骼的影响 - PTHRP中和抗体无法实现。 [D-Trp12，tyr34] -bovine-pth（7- 34），BPTH7-34DD，是pth1r的G蛋白偶联信号传导的独特反向激动剂，而cAMP降低了 激活β-arrestin信号传导。它会增加骨形成而不会引起高钙血症或骨溶解 [Maudsley等，2015]。我们的数据显示了BPTH7-34DD在共培养中的抗骨化和抗肿瘤作用 用小鼠骨骼乳腺癌。它还抵消了与癌症恶病质有关的脂肪细胞基因的变化。 我们假设BPTH7-34DD将：1）通过抑制骨化骨转移的恶性循环 溶解和肿瘤生长； 2）通过阻断白色到棕色脂肪细胞的转化来减少癌症恶病质 骨; 3）通过刺激新骨形成来保留骨骼健康； 4）有效治疗应有的骨质水解肿瘤 乳腺癌。 5）对其他PTHRP分泌类型有效。 BPTH7-34DD将在异种移植物中进行测试 乳腺癌骨转移的模型，预防和治疗环境。 BPTH7-34DD的机制 信号传导将在癌细胞的离体共培养中与小鼠骨骼以及骨骼和脂肪细胞系，以及 1O细胞以及来自WT＆β-arrestin k/o（βarr2 - / - ）小鼠的骨骼和细胞。 特定目标1：a）确定BPTH7-34DD对肿瘤细胞的生长效应； b）确定机制 i）成骨细胞（obs），破骨细胞（OCS）和骨细胞（OTS）的作用； ii）脂肪细胞； c）测试是否响应 对于BPTH7-34DD，使用βarr2 - / - 小鼠的骨骼和细胞需要β-arrestin； D）测试BPTH7-34DD是否阻止了增长 在骨骼和骨溶解中，具有前列腺和肺癌以及多发性骨髓瘤细胞的体内 特定目标2：在体内测试BPTH7-34DD，以实现以下方面的重大影响：a）肿瘤燃烧； b）溶性病变； c） 骨质流失/形成在肿瘤上且毗邻； D）骨骼肌。 意义：BPTH7-34DD可以迅速将其用于癌症骨转移的临床试验。类似的pth ＆PTHRP Pepperides用于治疗骨质疏松症。利益可能扩展到其他表达PTHRP的癌症 转移到骨头，包括多发性骨髓瘤，也将骨骼定居并引起溶解。 技术方法：该项目使用癌细胞与小鼠骨骼的共培养，称为Evoca（Ex vivo） 器官共培养分析）。肿瘤生长由稳定翻译的分泌的高斯荧光素酶（GLUC）分配 癌细胞。骨和MM基因的变化通过小鼠和人特异性QPCR进行定量。两只鼠标 乳腺癌骨转移模型将用于临床前测试。顾问：Louis Luttrell博士是 PTH信号的全球专家，并决定了BPTH7-34DD的行动。 Teresa Zimmers博士是专家 耶稣·德尔加多 - 卡尔博士是骨细胞生物学和骨细胞的专家：脂肪细胞 骨髓瘤的相互作用。戴维·罗德曼（David Roodman）博士是骨髓瘤骨病的世界专家。