Adipose-derived stem cell-conditioned medium therapy in a mouse model of ALS

脂肪干细胞条件培养基治疗 ALS 小鼠模型

• 批准号: 9921214
• 负责人: Chandler Walker
• 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 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9921214
• 关键词: ALS patients; Acute; Acute Lung Injury; Adipose tissue; Affect; American; Amyotrophic Lateral Sclerosis; Animal Model; Anti-Inflammatory Agents; Apoptosis; Apoptotic; Area; Axon; Biocompatible Materials; Biomedical Engineering; Blood Vessels; Blood specimen; Cardiovascular Pathology; Cell Culture Techniques; Cells; Cessation of life; Chronic; Collaborations; Conditioned Culture Media; Diagnosis; Disease; Disease Progression; Dose; Electromyography; Elements; Endothelium; Engineering; Experimental Animal Model; Gastrocnemius Muscle; Growth; Growth Factor; Gulf War; Hand Strength; Histologic; Histology; Hybrids; Hydrogels; Immune; Immune response; Immunologic Markers; Immunologics; In Vitro; Indiana; Individual; Inflammation; Inflammatory; Inflammatory Response; Injectable; Injections; Investigation; Ischemia; Kinetics; Light; Limb structure; Longevity; Lumbar spinal cord structure; Medial; Motor Activity; Motor Neurons; Mus; Muscle; Neuraxis; Neurodegenerative Disorders; Neuromuscular Junction; Onset of illness; Paralysed; Pathologic; Pathology; Patients; Peripheral; Persian Gulf; Phase I Clinical Trials; Polyethylene Glycols; Population; Quality of life; Research; Risk; Severity of illness; Site; Skeletal Muscle; Spinal Cord; Stromal Cells; Symptoms; Testing; Therapeutic; Therapeutic Effect; Time; Tissue Sample; Tissues; Treatment Efficacy; Treatment Protocols; United States; Universities; Veterans; clinical application; clinical candidate; clinical efficacy; clinical translation; crosslink; cytokine; design; effective therapy; experimental study; functional decline; functional loss; global health; immune activation; improved; in vivo; ineffective therapies; inflammatory marker; invention; loss of function; mouse model; neuromuscular; neuromuscular function; neuron loss; novel; novel therapeutics; preservation; protective factors; relating to nervous system; repaired; response; stem; stem cells; treatment effect

Amyotrophic lateral sclerosis (ALS) is a prevalent progressive neurodegenerative disease involving the loss of functional neuromuscular junctions (NMJ) and motor neurons (MN), and subsequent paralysis. No effective cures for ALS exist and current therapies only marginally impact disease course. This is because at the time of diagnosis, the disease has typically progressed into the final stages which impedes effective treatment. Therefore, treatment of ALS with novel therapies is an unmet need. Since veterans of the Persian Gulf War are twice as likely to develop ALS, discovering successful therapies is especially significant to our veteran population. Adipose-derived stem/stromal cells (ASC) have been shown to repair and rescue tissues from ischemia and other pathological conditions due to their beneficial secretions including anti-apoptotic, anti-inflammatory, and pro-angiogenic growth factors and cytokines. Recent studies have demonstrated the therapeutic value of conditioned medium isolated from ASC cultures (ASC-CM), and Phase I clinical trials utilizing ASC-CM to treat acute lung injury and specific cardiovascular pathologies are also ongoing. In this context, we have identified a B6SJL hybrid mSOD1G93A mouse model of ALS in which disease progression is similar to that observed in ALS patients. Evidence shows ASC-CM administration in this ALS mouse model after symptom onset improves lumbar spinal cord MN survival and prolongs lifespan. Moreover, early ASC-CM treatment (at disease onset) preserves intact NMJ. Therefore, early ALS treatment using ASC-CM may simultaneously target peripheral and central components of ALS. Using the mSOD1G93A ALS mouse model, we will also administer local muscular injection of a novel bioengineered ASC-CM-embedded polyethylene glycol (PEG) hydrogel that allows slow release of growth/protective factors at the site of neuromuscular disconnection to investigate the specific site(s) of ASC-CM action. We will also assess the overall efficacy for clinical translation of ASC-CM through optimized amelioration of disease progression. Three specific Aims have been designed to elaborate on this areas of investigation. Aim 1: Determine ASC-CM dose response on early NMJ loss, and assess neuromuscular and immune responses to long-term ASC-CM therapy in mSOD1G93A mice. A) First, an experiment will be performed to determine the optimal therapeutic dose of ASC-CM required to preserve intact NMJs. B) The optimal dose of ASC-CM from (A) will be administered from PD35-PD47 and NMJs and MN will be quantified weekly from PD56-91 and compared with C) the optimal dose of ASC-CM administered from PD35-continuously. Aim 2: Determine the therapeutic efficacy of early long-term continuous ASC-CM administration in mSOD1G93A mice. Using the optimal ASC-CM dose established in Aim 1, symptom onset, disease progression rate, and lifespan will be assessed and histologic analyses performed. To assess symptom onset and progression an array of functional assessments will be performed including electromyography (EMG), grip strength, locomotor activity, etc. Aim 3: Determine whether site of disease onset (NMJ) is the site of ASC-CM protective/reparative benefits using a novel slow-release biomaterial applied directly to muscle tissue in mSOD1G93A mice. Following in vitro assessment of ASC-CM release from PEG hydrogel, ASC-CM-PEG hydrogel will be injected into the medial gastrocnemius muscle and NMJ, MN loss and immune response over time will be examined. The results of this aim will shed light on the ASC-CM site of action on NMJ preservation and is a new clinically applicable invention for localized ASC-CM therapy.

肌萎缩侧索硬化症（ALS）是一种普遍存在的进行性神经退行性疾病，涉及 功能性神经肌肉接头（NMJ）和运动神经元（MN）的丧失，以及随后的 麻痹。 ALS 尚无有效治疗方法，目前的治疗方法只能对疾病产生轻微影响 课程。这是因为在诊断时，疾病通常已经发展到最终阶段 阻碍有效治疗的阶段。因此，采用新疗法治疗 ALS 是一种 未满足的需求。由于参加过波斯湾战争的退伍军人患 ALS 的可能性是其两倍，因此发现 成功的治疗对于我们的退伍军人群体尤其重要。脂肪来源 干/基质细胞 (ASC) 已被证明可以修复和拯救缺血和其他疾病的组织 由于其有益的分泌物而产生的病理状况，包括抗凋亡、抗炎、 以及促血管生成生长因子和细胞因子。最近的研究表明，治疗 从 ASC 培养物中分离的条件培养基 (ASC-CM) 的价值和 I 期临床试验 利用 ASC-CM 治疗急性肺损伤和特定心血管疾病也在进行中。 在此背景下，我们鉴定了 ALS 的 B6SJL 杂交 mSOD1G93A 小鼠模型，其中疾病 进展与 ALS 患者中观察到的相似。有证据表明 ASC-CM 给药 这种 ALS 小鼠模型在症状出现后改善了腰脊髓 MN 的存活并延长了时间 寿命。此外，早期 ASC-CM 治疗（疾病发作时）可保留完整的 NMJ。所以， 使用 ASC-CM 进行早期 ALS 治疗可能同时针对外周和中枢部位 肌萎缩侧索硬化症。使用 mSOD1G93A ALS 小鼠模型，我们还将进行局部肌肉注射 一种新型生物工程 ASC-CM 嵌入聚乙二醇 (PEG) 水凝胶，可缓慢 在神经肌肉断开部位释放生长/保护因子以研究 ASC-CM 作用的特定位点。我们还将评估临床转化的总体疗效 ASC-CM 通过优化改善疾病进展。制定了三个具体目标 旨在详细说明这一调查领域。 目标 1：确定 ASC-CM 对早期 NMJ 损失的剂量反应，并评估神经肌肉和 mSOD1G93A 小鼠对长期 ASC-CM 治疗的免疫反应。 A) 首先，进行一个实验 以确定保存完整 NMJ 所需的 ASC-CM 最佳治疗剂量。二） (A) 中的 ASC-CM 的最佳剂量将从 PD35-PD47 中施用，并且 NMJ 和 MN 将是 每周从 PD56-91 定量，并与 C) 相比，ASC-CM 的最佳剂量从 PD35-连续。 目标 2：确定早期长期连续 ASC-CM 给药的治疗效果 在 mSOD1G93A 小鼠中。使用目标 1、症状出现、疾病中确定的最佳 ASC-CM 剂量 将评估进展率和寿命并进行组织学分析。评估症状 发病和进展将进行一系列功能评估，包括肌电图 （肌电图）、握力、运动活动等。 目标 3：确定疾病发作部位 (NMJ) 是否是 ASC-CM 保护/修复部位 使用直接应用于 mSOD1G93A 肌肉组织的新型缓释生物材料的好处 老鼠。对 PEG 水凝胶中 ASC-CM 的释放进行体外评估后，ASC-CM-PEG 水凝胶将 注射到腓肠肌内侧，随着时间的推移，NMJ、MN 损失和免疫反应 将接受检查。这一目标的结果将揭示 ASC-CM 在 NMJ 保护方面的作用点 是一项临床适用的局部ASC-CM治疗的新发明。