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: 10582528
• 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/10582528
• 关键词: ALS pathology; 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; Cells; Central Nervous System; 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; Neurodegenerative Disorders; Neuromuscular Junction; Onset of illness; Paralysed; Pathologic; Pathology; Patients; Peripheral; Persian Gulf; Phase I Clinical Trials; Polyethylene Glycols; 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; adipose derived stem cell; amyotrophic lateral sclerosis therapy; 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; military veteran; mouse model; neural; neuromuscular; neuromuscular function; neuron loss; neuronal survival; neuroprotection; novel; novel therapeutics; preservation; protective factors; repaired; response; stem; 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.

肌萎缩性侧索硬化症（ALS）是一种普遍的进行性神经退行性疾病，涉及 功能性神经肌肉连接（NMJ）和运动神经元（MN）的丧失以及随后的 麻痹。没有有效的ALS治疗方法，当前疗法仅对疾病产生少量影响 课程。这是因为在诊断时，该疾病通常已经发展为最终 阻碍有效治疗的阶段。因此，使用新型疗法的ALS治疗是一种 未满足的需求。由于波斯湾战争的退伍军人发展ALS的可能性是 成功的疗法对我们的资深人口尤为重要。脂肪来源 茎/基质细胞（ASC）已显示可修复和营救缺血和其他的组织 病理条件由于其有益分泌物，包括抗凋亡，抗炎， 和促血管生长因子和细胞因子。最近的研究证明了治疗性 从ASC培养物（ASC-CM）和I期临床试验中分离出的条件培养基的值 利用ASC-CM治疗急性肺损伤和特定的心血管病理也正在进行中。 在这种情况下，我们确定了B6SJL混合MSOD1G93A ALS的小鼠模型，其中 进展与ALS患者观察到的进展相似。有证据表明ASC-CM给药 症状发作后，这种ALS小鼠模型改善了腰椎MN存活并延长 寿命。此外，早期ASC-CM治疗（在疾病发病时）可保留完整的NMJ。所以， 使用ASC-CM的早期ALS处理可能同时靶向外围和中央组件 ALS。使用MSOD1G93A ALS小鼠模型，我们还将管理局部肌肉注射 一种新型的生物工程ASC-CM包裹的聚乙二醇（PEG）水凝胶，可慢慢 在神经肌肉断开的部位释放生长/保护因子以研究 ASC-CM动作的特定部位。我们还将评估用于临床翻译的总体功效 ASC-CM通过优化疾病进展的改善。三个具体目标是 旨在详细说明这一调查领域。 目标1：确定对早期NMJ损失的ASC-CM剂量反应，并评估神经肌肉和 MSOD1G93A小鼠中对长期ASC-CM治疗的免疫反应。 a）首先，实验将 进行以确定保留完整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PD35-PD47和NMJ和MN施用 每周从PD56-91进行量化，并与C）进行比较 PD35连续。 目标2：确定早期连续ASC-CM给药的治疗功效 在MSOD1G93A小鼠中。使用AIM 1中建立的最佳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 发作和进展将进行一系列功能评估，包括肌电图 （EMG），抓地力，运动活性等。 AIM 3：确定疾病发作部位（NMJ）是否是ASC-CM保护/修复的部位 使用新型的慢透明生物材料直接应用于MSOD1G93A中的肌肉组织的好处 老鼠。遵循体外评估PEG水凝胶释放的ASC-CM释放，ASC-CM-PEG水凝胶将 随着时间的流逝，被注射到内侧胃gus肌和NMJ，MN损失和免疫反应 将被检查。此目标的结果将揭示在NMJ保存的ASC-CM动作部位 它是一种用于局部ASC-CM治疗的新临床适用发明。

项目成果

Progress in perisynaptic Schwann cell and neuromuscular junction research.

• DOI: 10.4103/1673-5374.327334
• 发表时间: 2022-06
• 期刊: Neural regeneration research
• 影响因子: 6.1
• 作者: Walker CL

Adipose-derived stem cell conditioned medium for the treatment of amyotrophic lateral sclerosis: pre-clinical evidence and potential for clinical application.

脂肪干细胞条件培养基用于治疗肌萎缩侧索硬化症：临床前证据和临床应用潜力。

• DOI: 10.4103/1673-5374.253514
• 发表时间: 2019
• 期刊: Neural regeneration research
• 影响因子: 6.1
• 作者: Walker,ChandlerL

Bisperoxovanadium promotes motor neuron survival and neuromuscular innervation in amyotrophic lateral sclerosis.

• DOI: 10.1186/s13041-021-00867-7
• 发表时间: 2021-10-11
• 期刊: Molecular brain
• 影响因子: 3.6
• 作者: Wang J;Tierney L;Mann R;Lonsway T;Walker CL
• 通讯作者: Walker CL

Carboxyl-terminal modulator protein (CTMP) deficiency mitigates denervation-induced skeletal muscle atrophy.

• DOI: 10.1016/j.bbrc.2023.01.023
• 发表时间: 2023-01
• 期刊: Biochemical and biophysical research communications
• 影响因子: 3.1
• 作者: Junmei Wang;Lydia Tierney;Christopher Wilson;V. Phillips;Lillian Goldman;Christen L. Mumaw;En Muang-En