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， 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 瘫痪ALS没有有效的治疗方法，目前的治疗方法对疾病的影响很小 当然了这是因为在诊断时，疾病通常已进展到最终阶段。 阻碍有效治疗的阶段。因此，用新疗法治疗ALS是一种有效的方法。 未满足的需求由于波斯湾战争的退伍军人患肌萎缩侧索硬化症的可能性是常人的两倍， 成功的治疗对我们的退伍军人群体尤其重要。脂肪来源 干/基质细胞（ASC）已经显示出修复和拯救组织免于缺血和其它损伤， 由于它们的有益分泌物，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， 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的心血管病变也在进行中。 在此背景下，我们已经鉴定了ALS的B6 SJL杂交mSOD 1G 93 A小鼠模型，其中疾病 进展与ALS患者中观察到的相似。证据显示ASC-CM给药 这种ALS小鼠模型在症状发作后改善了腰髓MN存活和存活率。 寿命此外，早期ASC-CM治疗（在疾病发作时）保留了完整的NMJ。因此，我们认为， 使用ASC-CM的早期ALS治疗可以同时靶向外周和中枢成分 的ALS。使用mSOD 1G 93 A ALS小鼠模型，我们还将局部肌肉注射 一种新型生物工程ASC-CM-嵌入的聚乙二醇（PEG）水凝胶， 在神经肌肉切断部位释放生长/保护因子，以研究 ASC-CM作用的特定部位。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。三个具体目标是 旨在详细阐述这方面的调查。 目的1：确定ASC-CM对早期NMJ损失的剂量反应，并评估神经肌肉和 mSOD 1G 93 A小鼠中对长期ASC-CM治疗的免疫应答。A.首先，一个实验将 以确定保存完整NMJ所需的ASC-CM的最佳治疗剂量。（B） 来自（A）的ASC-CM的最佳剂量将从PD 35-PD 47施用，并且NMJ和MN将从PD 35-PD 47施用。 从PD 56 -91每周定量，并与C）从PD 56 - 91每周定量施用的ASC-CM的最佳剂量进行比较。 PD 35-连续。 目的2：确定早期长期连续ASC-CM给药的治疗效果 在mSOD 1G 93 A小鼠中。使用目标1中确定的最佳ASC-CM剂量，症状发作，疾病 将评估进展率和寿命，并进行组织学分析。评估症状 将进行一系列功能评估，包括肌电图检查 (EMG)握力、自发活动等。 目的3：确定疾病发作部位（NMJ）是否为ASC-CM保护/修复部位 在mSOD 1G 93 A中使用直接应用于肌肉组织的新型缓释生物材料的益处 小鼠在体外评估ASC-CM从PEG水凝胶的释放后，ASC-CM-PEG水凝胶将 注射到腓肠肌内侧和NMJ，MN损失和免疫反应随着时间的推移 将被审查。这一目标的结果将阐明ASC-CM对NMJ保存的作用位点 是ASC-CM局部治疗的临床应用新发明。