本發明提供用於幹細胞療法中，用於治療polyQ疾病之疾病或病況的方法及製品。polyQ疾病係由各別蛋白質中編碼長polyQ鏈之胞嘧啶-腺嘌呤-鳥嘌呤(CAG)重複序列擴增引起的一組神經退化性病症。PolyQ疾病係以不相關基因之轉譯區中CAG三核苷酸重複之病理性擴增為特徵。經轉譯之polyQ聚集在退化之神經元內，導致特定神經元亞群之功能障礙及退化。本發明出人意料地發現利用幹細胞之治療方案，該治療方案提供用於恢復polyQ疾病中退化及/或受損神經元之功能的有效療法。 除非另外指出，否則技術性術語係根據習知用法使用。 如本文中所使用，單數形式「一個(種)(a/an)」及「該/該等」係指單數以及複數兩者，除非上下文另有清晰指示。 如本文所使用，術語「及」及「或」可用於指連接性或分離性的。亦即，該兩個術語應理解為等效於「及/或」，除非另有說明。 如本文所用，術語「治療(treatment)」、「治療(treat)」或「治療(treating)」係指疾病或病況或病症，或症狀、不良作用或後果，或與之相關之表型的完全或部分改善或減輕。希望的治療作用包括(但不限於)防止疾病之發生或復發、緩解症狀、減少疾病之任何直接或間接病理性後果、降低疾病進展速率及改善減輕疾病狀態。 如本文所用，術語「延緩疾病發展」意指推遲、阻礙、減慢、停滯、穩定、抑制及/或延遲疾病之發展。取決於所治療之疾病及/或個體之病史，此延緩可具有不同的時間長度。 如本文所用，在投藥之情況下，術語藥劑(例如醫藥調配物、細胞或組合物)之「有效量」係指以所需劑量/量且持續所需時段有效獲得所希望之結果的量。 如本文所用，術語藥劑(例如醫藥調配物或細胞)之「治療有效量」係指以所需劑量且持續所需時段有效獲得所希望之治療結果(諸如用於治療疾病、病況或病症，及/或治療之藥物動力學或藥效學作用)的量。 如本文所用，「第一次劑量」係用於描述給定劑量係在投與連續或後續劑量之前的時序。該術語未必暗示個體之前從未接受過一劑細胞療法或甚至個體之前從未接受過一劑相同細胞。 如本文所用，術語「後續劑量」係指在前一次劑量(例如，第一次劑量)之後向同一個體投與的劑量，在此期間未向個體投與任何插入劑量。 如本文所用，術語「個體」係哺乳動物，諸如人類或其他動物，且通常為人類。在一些實施例中，個體在投藥之前曾利用靶向疾病或病況之治療劑治療。 如本文所用，術語「醫藥調配物」係指這樣一種製劑，其形式允許其中所含活性成分之生物活性有效，且其不含對投與該調配物之個體具有不可接受之毒性的額外組分。 如本文所用，術語「醫藥學上可接受之載劑」係指醫藥調配物中除活性成份以外的對個體無毒之成份。醫藥學上可接受之載劑包括(但不限於)緩衝劑、賦形劑、穩定劑或防腐劑。 在一個態樣中，本發明提供一種用於治療個體之多麩醯胺酸(polyQ)疾病的方法，該方法包含向個體非經腸或局部地投與有效量之幹細胞作為單位劑量，其中該投與係以一或多個治療週期進行，其中一個治療週期包含分別以2至6週的給藥間隔給與三個單位劑量。 在一些實施例中，polyQ疾病包括(但不限於)脊髓小腦性共濟失調(SCA)；馬查多-約瑟夫病(MJD/SCA3)；亨廷頓氏病(HD)；齒狀紅核蒼白球路易體萎縮(DRPLA)；及1型X連鎖脊髓延髓性肌肉萎縮(SMAX1/SBMA)。 在一些實施例中，SCA係多麩醯胺酸(polyQ)介導之SCA，SCA較佳為SCA1、SCA2、SCA3、SCA6、SCA7或SCA17。更佳地，SCA為SCA3。 在一些實施例中，間葉幹細胞係間葉幹細胞群(MSC)、脂肪組織源性幹細胞(ADMSC)群、眼眶脂肪源性幹細胞(OFSC)群或四潛能陽性基質細胞(QPSC)群。在一個實施例中，QPSC係描述於美國申請案第14/615,737號之QPSC，其具有至少70%細胞同源性且表現細胞標記物CD273、CD46、CD55及CXCR4，但不表現CD45；其中CD273以超過70%之強度較強地表現。在一個實施例中，ADSC係描述於美國20120288480中之彼等OFSC，其至少表現CD90、CD 105、CD29、CD44、CD49b、CD49e、CD58及HLA-ABC，但不表現CD133、CD31、CD106、CD146、CD45、CD14、CD117。幹細胞較佳為QPSC群。 在一些實施例中，細胞可藉由非經腸投與或局部治療投與(諸如腦內或顱內投與)。非經腸輸注包括肌內、靜脈內、動脈內或皮下投與。非經腸投與較佳為靜脈內注射。 在一些實施例中，單位劑量在0.5×105
至5×1010
個細胞/kg體重範圍內。在一些實施例中，單位劑量在0.5×105
至5×109
、0.5×105
至5×108
、0.5×105
至5×107
、0.5×105
至5×106
、1.0×105
至5×1010
、1.0×105
至5×109
、1.0×105
至5×108
、1.0×105
至5×107
或1.0×105
至5×106
個細胞/kg體重範圍內。 在一個實施例中，投與係以一或多個治療週期進行，其中一個治療週期包含分別以2至6週(亦即，兩週、三週、四週、五週或六週。在另一實施例中，間隔為兩週)的給藥間隔給與三個單位劑量。本發明之治療週期的數目係根據共濟失調評估及分級量表(scale for the assessment and rating of ataxia，SARA)確定(Subramony SH., SARA--a new clinical scale for the assessment and rating of ataxia. Nat Clin Pract Neurol. 2007; 3(3):136-7；Kim BR, Lim JH, Lee S, Park S, Koh SE, Lee IS, Jung H, Lee J. Usefulness of the Scale for the Assessment and Rating of Ataxia (SARA) in Ataxic Stroke Patients.Ann Rehabil Med
. 2011; 35: 772-780；及Tan S, Niu HX, Zhao L等人, Reliability and validity of the Chinese version of the Scale for Assessment and Rating of Ataxia.Chin Med J
. 2013; 126(11):2045-8)。SARA係基於小腦共濟失調(脊髓小腦，弗里德希氏(Friedreich)及散發性共濟失調)損傷水準之半定量評估的臨床量表。SARA係基於8個項目之表現的量表，得到總分0分(無共濟失調)至40分(最嚴重共濟失調)。該等分數係基於步態、跨距、坐姿、言語障礙、手指追蹤(finger chase)、鼻-手指測試、快速交替手運動及踵-脛滑動的患者表現。在第一個治療週期之後，若個體保持SARA總分高於5分一個月，則將進行第二個及後續治療週期。 單位劑量之幹細胞係以2至6週之間隔投與。2至6週之間隔意指單位劑量之幹細胞係在兩週、三週、四週、五週或六週內投與一次。在一個實施例中，給藥間隔係每兩週一次。在一個實施例中，每兩週一次給藥意指單位劑量之幹細胞係兩週一次投與，亦即，在14天時段期間投與一次，較佳在每兩週的同一天投與。在每兩週一次給藥方案中，大體上約每14天投與單位劑量。 在幹細胞療法情況下，單位劑量之投與包含以單一組合物及/或單次不間斷的投藥(例如，單次注射或連續輸注)投與給定量或數目之細胞。 在一些實施例中，細胞係作為組合治療之一部分，諸如與另一治療干預同時或按任何次序依序投與。在一些實施例中，幹細胞係與一或多種額外治療劑或結合另一治療干預同時或按任何次序依序共投與。在一些情況下，該等額外治療劑或另一療法係與細胞以足夠接近的時間共投與，由此使該等額外治療劑或另一療法增強細胞群的作用，或反之亦然。在一些實施例中，幹細胞係在該一或多種額外治療劑之前投與。在一些實施例中，幹細胞係在該一或多種額外治療劑之後投與。 用於本發明方法中之幹細胞係調配為醫藥組合物或調配物形式，諸如包括供投與給定劑量或其部分之細胞數目的單位劑型組合物。醫藥組合物及調配物一般包括一或多種可選的醫藥學上可接受之載劑或賦形劑。在一些實施例中，該組合物包括至少一種額外治療劑。 載劑之選擇部分由特定幹細胞及/或由投與方法確定。因此，存在多種適合調配物。舉例而言，該醫藥組合物可含有防腐劑。 醫藥學上可接受載劑以所用劑量及濃度大體上對接受者無毒，且包括(但不限於)：緩衝劑，諸如磷酸鹽、檸檬酸鹽及其他有機酸；抗氧化劑，包括抗壞血酸及甲硫胺酸；防腐劑(諸如氯化十八烷基二甲基苯甲基銨；氯化六羥季銨；苯紮氯銨(benzalkonium chloride)；苄索氯銨(benzethonium chloride)；苯酚、丁醇或苄醇；對羥基苯甲酸烷酯，諸如對羥基苯甲酸甲酯或對羥基苯甲酸丙酯；兒茶酚；間苯二酚；環己醇；3-戊醇；及間甲酚)；低分子量多肽；蛋白質，諸如血清白蛋白、明膠或免疫球蛋白；親水性聚合物，諸如聚乙烯吡咯啶酮；胺基酸，諸如甘胺酸、麩醯胺酸、天冬醯胺、組胺酸、精胺酸或離胺酸；單醣、雙醣，及其他碳水化合物，包括葡萄糖、甘露糖或糊精；螯合劑，諸如EDTA；糖，諸如蔗糖、甘露醇、海藻糖或山梨醇；成鹽相對離子，諸如鈉；金屬錯合物(例如，Zn-蛋白質錯合物)；及/或非離子界面活性劑，諸如聚乙二醇(PEG)。 在一些態樣中，緩衝劑包括在組合物中。適合的緩衝劑包括例如檸檬酸、檸檬酸鈉、磷酸、磷酸鉀，以及各種其他酸及鹽。在一些態樣中，使用兩種或多於兩種緩衝劑之混合物。製備可投與之醫藥組合物的方法係已知的。 調配物可包括水溶液。調配物或組合物亦可含有多於一種可用於以幹細胞治療之特定適應症、疾病或病況的活性成份。 在一些實施例中，醫藥組合物包含有效治療疾病或病況之量(諸如治療有效量)的幹細胞。在一些實施例中，藉由定期評估所治療之個體來監測治療功效。所需劑量可藉由單次快速注射投與細胞、藉由多次快速注射投與細胞或藉由連續輸注投與細胞來遞送。 幹細胞及組合物可使用標準投藥技術、調配物及/或裝置投與。幹細胞之投與可為自體或異源的。 無菌可注射溶液可藉由將細胞併入溶劑中，諸如使細胞與適合之載劑、稀釋劑或賦形劑(諸如無菌水、生理食鹽水、葡萄糖、右旋糖或其類似物)混合來製備。取決於投藥途徑及所需製劑，組合物可含有輔助物質，諸如潤濕劑、分散劑或乳化劑(例如甲基纖維素)、pH緩衝劑、膠凝或黏度增強添加劑、防腐劑、調味劑及/或著色劑。在一些態樣中，可參考標準文本以製備適合製劑。 可添加增強組合物之穩定性及無菌性的各種添加劑，包括抗微生物防腐劑、抗氧化劑、螯合劑及緩衝劑。可藉由各種抗細菌劑及抗真菌劑(例如，對羥基苯甲酸酯、氯丁醇、苯酚及山梨酸)來確保防止微生物之作用。可藉由使用吸收延遲劑(諸如單硬脂酸鋁及明膠)來延長可注射醫藥形式之吸收。實例 I. 動物模型試驗 材料及方法 動物及實驗設計 無顯著運動功能退化之小鼠
MJD84.2(B6;CBA-Tg(ATXN3*)84.2Cce/IbezJ)小鼠已確立為人類馬查多-約瑟夫疾病，亦稱為3型脊髓小腦性共濟失調(MJD/SCA3)之疾病模型。在本研究中，研究年齡在20至34週範圍內的MJD84.2動物。對此等動物進行行為分析，包括改良之SHIRPA、腳印分析及旋桿測試。以兩週間隔對21、23及25週齡小鼠進行三次測試物注射。研究設計概述如下。 具有顯著運動功能退化之小鼠
總計十三隻SCA3 Tg/0小鼠(B6;CBA-Tg (ATXN3*)84.2Cce/IbezJ)及8隻C57BL/6 0/0野生型小鼠最初全部來自JAX實驗室。動物隨機登記分入四個實驗組：(1) SCA3+細胞；(2) SCA3+PBS；(3) Wt+細胞；(4) Wt+PBS。在確定SCA3 Tg/0小鼠出現明顯著疾病表型之後以兩週間隔進行三次測試物注射。研究設計概述如下。 間葉幹細胞
本研究中之QPSC為人類ADMSC (Stemchymal®)，即由Steminent Biotherapeutics Inc. (SBI)製造的細胞產品。在根據PIC/S良好工廠實踐指南(Good Manufactory Practice guideline)所建立之細胞工廠中，遵循SBI標準操作程序，對ADMSC進行離體培養擴增及品質控制。簡言之，自健康供體收集脂肪組織且立即在低溫下(0-5℃)運送至SBI加工設施中。分離ADMSC，純化且在培養物擴增期間，保持在SBI獨有的培養基中。第12代ADMSC係高水準表現CD273、CD46、CD55及CXCR4的QPSC，隨後將其包裝於低溫保藏袋中，且產品(Stemchymal®)送交品質檢定及低溫保藏。Stemchymal®之品質控制係由製程內控制及產物放行測試組成，其包括(但不限於)存活性、無菌性、黴漿菌測試、內毒素評估、MSC表型(針對CD 73、CD 90及CD 105呈陽性，針對CD 34、CD 45、CD11b、CD 19及HLA-DR呈陰性)及三譜系分化能力(成骨分化、成軟骨分化及成脂肪分化)。 細胞給與 對於使用無顯著運動功能退化之小鼠的研究 ：
將2.5×107
個細胞/kg體重之Stemchymal®
解凍，準備好並裝載入1 ml胰島素注射器(29 1/2G)中。在解凍一小時之內緩慢注射細胞(15至20秒的持續時間)。對於使用具有顯著運動功能退化之小鼠的研究 ：
小鼠隨機分入四個組：(1) SCA3+細胞；(2) SCA3+PBS；(3) Wt+細胞；(4) Wt+PBS。將2.5×107
個細胞/kg體重之Stemchymal®
靜脈內輸注至第1組及第3組中的每隻小鼠。向各小鼠投與總計125 μl之細胞懸浮液(1:1低溫溶液(Biolife))或PBS(Gibco)。 對於該兩項研究，在注射之後監測動物4個小時且每日觀測。每兩週一次給與細胞，總計三次。 數據收集 及 分析
在最後一次測試物注射之後一個月處死小鼠。記錄整個研究中小鼠之體重及掉落之等待時間(latency to fall)。在測試物注射之後，亦針對步態表現分析小鼠腳印。收集小鼠組織(皮層、小腦、心臟、腎臟、肝、脾、肺及尾)以用於將來組織病理學分析及生物學分佈研究。 統計學
數據以平均值±SEM呈現。使用斯圖登氏t檢驗(Student's t-test)且以p＜0.05之顯著性臨限值分析旋桿及腳印測試之結果。 運動協調及平衡分析
在旋桿設備(MK-670, Muromachi Kikai Co., Ltd., Japan)中評價運動協調及平衡情況。將小鼠置放於處於恆定速度(4 rpm)之旋桿上，該旋桿在5 min時段內加速至40 rpm。記錄掉落之等待時間或者完成完整被動旋轉(附著至桿上完整旋轉)之等待時間。使小鼠進行3次各測試，其中各試驗之間有15 min休息。計算各測試中各小鼠之平均等待時間。測試結果使用t檢驗進行統計分析。 SHIRPA 測試
改良之SHIRPA測試係在小鼠年齡為20、24及28週時進行。SHIRPA方案係改自RIKEN BRC之改良之SHIRA方案。測試項目及評分標準列於下表中。
小鼠係根據其行為評分。研究組中小鼠總數計為100%。結果以某些評分層級之小鼠數目的百分比呈現。 腳印分析
在最後一次給與細胞之後約一個月分析小鼠腳印。參考2015(1)所公佈的論文，將小鼠腳掌浸漬於墨水中(前腳：紅色；後腳：綠色)，以使得小鼠在沿走廊步行或跑至目標箱時留下腳印的形跡。將將小鼠置放於隧道前方的一張紙(50 cm長，10 cm寬)上。量測步幅、搖擺、跨距長度以及前腳爪及後腳爪重疊指示步態(參看下圖)。所有小鼠在處死之前進行三輪量測。 MTT 分析 CD3 + T 細胞分離
藉由Histopaque-1077(Sigma-Aldrich)密度梯度離心由來自健康供體之肝素化全血分離人類末梢血液單核細胞(PBMC)。隨後，遵循製造商的說明，使用抗人類CD3抗體偶聯之磁性粒子(BD Biosciences)藉由陽性選擇自PBMC純化出CD3+
T淋巴球。T 細胞 增殖分析
在96孔盤中，利用盤結合之抗-CD3單株抗體(2 μg/ml)及抗-CD28單株抗體(2 μg/ml) (BD Biosciences)刺激經純化之人類CD3+
T細胞(1×105
個細胞)且將其與不同數量之ADMSC共培養於含有10%胎牛血清(FBS)、2 mM l-麩醯胺酸、100 U/ml青黴素、100 U/ml鏈黴素及25 mM HEPES RPMI-1640培養基(Gibco)中。在48小時之後，將5-溴-2-脫氧尿苷(BrdU)添加至各孔中且再培育盤18小時以量測T細胞增殖情況。根據製造商之說明，使用細胞增殖ELISA(Cell Proliferation ELISA) BrdU套組(Roche)量測併入T細胞中之BrdU之量。 免疫組織化學 ( IHC )
為評價QPSC之神經保護作用，將QPSC經由C57BL/6J SCA2轉殖基因小鼠之尾靜脈(IV hMSC-Tg組)或經由枕骨大孔(IC hMSC-Tg組)注射入小腦位置。選擇與人類β2微球蛋白反應之特異性抗體(Abcam，代碼：ab15976)，藉由IHC展示鼠類腦組織中之人類細胞。切割鼠類切片(4 μm)並安放至顯微鏡載片上。藉由在二甲苯、100%乙醇、95%乙醇及80%乙醇中以5 min間隔沖洗兩次使切片復水。在去除石蠟之後，切片用3% H2
O2
處理以使過氧化酶失活，在10 mM檸檬酸鹽緩衝液(含0.05% Tween20)中加熱以用於抗原修復，且用1%阻斷溶液阻斷(1% BSA及0.1% Triton X-100(於PBS中)，Chang等人，Journal of Biomedical Science 2011, 18:54 http://www.jbiomedsci.com/content/18/1/54，第3頁，共9頁)。將切片與在阻斷溶液中稀釋(1:400)之特異性抗人類b2微球蛋白多株抗體(Abcam)一起在室溫下培育40 min。在用PBS充分洗滌三次之後，將切片與在阻斷溶液中稀釋(1:1000)之二次抗體一起在室溫下培育40 min。一次抗體使用EnVision偵測系統(DAKO)偵測，且用二胺基聯苯胺(DAB；DAKO)觀測。用含水蘇木精(SigmaAldrich)對比染色。對於直接比較，整批處理所有載片以使變異性最低。 安全性測試 動物
C57BL/6小鼠接受使用胰島素注射器1/2cc 30G x 3/8''針頭(Terumo，或BD Bioscience)經由尾部靜脈內注射的3劑QPSC。在注射之前，動物藉由置放在籠子下的加熱墊溫熱15至20分鐘以擴張其尾靜脈。在屍檢之前，所有動物使用胺基甲酸酯(2公克/公斤體重，Sigma-Aldrich)麻醉，接著自下頜下靜脈或經由心臟穿刺收集血液。血液樣品收集
對於血液學分析，將全血樣品收集在含有EDTA之採血管(BD Bioscience, 目錄號365974)中。對於血液化學分析，將全血樣品收集在含有血漿分離器之採血管(BD Bioscience, 目錄號365967)中。在室溫下靜置血漿管20 min之後，接著藉由在4℃下以6000rpm離心5 min分離血漿。肉眼屍檢及組織收集
在取血樣之後，收集動物器官。將其各分成兩部分：(1)一半器官保藏於-80℃冷凍器中，隨後轉移且儲存在液氮容器中以用於生物分佈分析；(2)另一半經固定(4%多聚甲醛，Sigma-Aldrich)且石蠟包埋以用於組織病理學分析。 定量 PCR
遵循製造商的說明，使用總RNA小規模純化套組(Total RNA Miniprep Purification Kit；GMbiolab目錄號TR01)自QPSC或鼠類組織提取總RNA。隨後，使用兩步MMLV RT-PCR套組(GMbiolab目錄號RP012-M)進行cDNA合成。使用The Fast SYBR® Green Master Mix (Thermo目錄號4385612)進行定量PCR以分析所選基因之相對表現。 ELISA
為量測QPSC中EGF、FGF-b、VEGF、PDGF及TGF-b1之細胞內及分泌含量，如下製備細胞溶解產物及改良性培養基之樣品：藉由使用凍融方法溶解QPSC，且在超速離心之後收集細胞溶解產物之上清液。且對於改良性培養基收集，在QPSC培養3天之後收集培養基。最後，根據製造商的說明，藉由ELISA(R&D system)測定以上生長因子之濃度。 神經元細胞共培養測試
人類星形膠質細胞株SVG p12用1250 μM 1-甲基-4-苯基吡啶(MPP+)處理且與不同比例之QPSC(SVG p12: QPSC=1:0.1-1:10)共培養。在24小時之後，計算SVG p12的細胞數目。實例 1 QPSC 改變 SCA3 之表型
QPSC改變SCA3小鼠之表型。如圖1所示，SCA3小鼠展示與野生型小鼠相比略微較寬的基部(base)且在QPSC治療之後，SCA3小鼠的外觀看起來類似於野生型小鼠。在各種功能測試中，諸如改良之SHIRPA(圖4、圖5)、腳印(圖6、圖7)及旋桿表現分析(圖5C)，觀測到類似的改善結果。實例 2 藉由本發明之治療使體重減輕停止且對器官組織無副作用
QPSC亦停止在疾病進展期間SCA3之體重減輕(圖3)。儘管如此，3次劑量之QPSC並不影響SCA3個體之全血球計數(表1)或血液生物化學(表2)的型態。表1及表2展示25至30週齡野生型小鼠的全血球計數/生物化學型態在三次劑量之QPSC與生理食鹽水(以一週間隔投與三次劑量)之間並無差異。組織病理學分析展示在注射三次劑量之QPSC之後各種重要器官組織的正常發現(圖2)。 表1
數據以平均值±SD呈現 表2
數據以平均值±SD呈現實例 3 使用本發明治療之小鼠中的免疫調節及抗 ROS 能力以及多個神經營養因子及生長因子之表現
活體外研究展示，QPSC不僅具有免疫調節及抗ROS能力(圖10)，而且亦具有表現多個神經營養因子及生長因子的能力(圖12)。活體內研究表明在QPSC治療之後，SCA小鼠在氧化應力下旋桿表現改善(圖11)。此外，QPSC亦可同時在活體外(圖13)及活體內(圖14)防止神經元損失。儘管已提出有關細胞跨血腦屏障(BBB)遷移之可能性的疑問，但已顯示QPSC經由靜脈內輸注而在顱內定位的能力(圖8及圖9)。 因此，合理地得出以下結論：經由靜脈內輸注，QPSC可通過BBB到達小腦且保護SCA個體之神經元細胞免受ROS及免疫過度反應的損害。QPSC亦分泌多個神經營養及生長因子以維持神經元細胞的數目，從而延緩poly-Q疾病(諸如多麩醯胺酸脊髓小腦性共濟失調、馬查多-約瑟夫疾病、亨廷頓氏病、DRPLA及SMAX1/SBMA)的進展。II. 人類臨床試驗
該人類臨床試驗旨在藉由隨機化、雙盲、安慰劑對照研究設計來研究Stemchymal®輸注用於治療多麩醯胺酸介導之疾病(諸如，多麩醯胺酸脊髓小腦性共濟失調、馬查多-約瑟夫疾病、亨廷頓氏病、DRPLA及SMAX1/SBMA)之治療功效及安全性。符合條件的個體將接受經由靜脈內輸注的Stemchymal®。 在針對多麩醯胺酸脊髓小腦性共濟失調的一個實例中，經歷試驗之個體具有經基因型確認的2型脊髓小腦性共濟失調或3型脊髓小腦性共濟失調。個體的基線SARA分數在5至15分範圍內。 將2.5×107
個細胞/kg體重之Stemchymal®
解凍、準備好並裝載入注射器中。在解凍一小時之內緩慢注射細胞。將Stemchymal®
經靜脈內輸注給各個體且以每兩週一次之間隔進行3次細胞給藥。在一或多個治療週期之後，個體之SARA分數降低且SCA2或SCA3病況改善。The present invention provides methods and articles of manufacture for the treatment of diseases or conditions of polyQ diseases in stem cell therapy. The polyQ disease is a group of neurodegenerative disorders caused by amplification of cytosine-adenine-guanine (CAG) repeats encoding long polyQ chains in individual proteins. PolyQ disease is characterized by pathological expansion of CAG trinucleotide repeats in translational regions of unrelated genes. The translated polyQ is concentrated in degenerating neurons, resulting in dysfunction and degradation of specific neuronal subpopulations. The present invention surprisingly finds a therapeutic regimen utilizing stem cells that provides an effective therapy for restoring the function of degenerating and/or damaged neurons in polyQ disease. Unless otherwise indicated, technical terms are used according to conventional usage. The singular forms "a", "the" and "the" and "the" As used herein, the terms "and" and "or" may be used to refer to the <RTI ID=0.0> That is, the two terms are to be understood as equivalent to "and/or" unless otherwise stated. As used herein, the terms "treatment", "treat" or "treating" refer to a disease or condition or disorder, or a symptom, an adverse effect or consequence, or a complete phenotype associated therewith. Or partially improved or mitigated. Desirable therapeutic effects include, but are not limited to, preventing the onset or recurrence of the disease, alleviating symptoms, reducing any direct or indirect pathological consequences of the disease, reducing the rate of disease progression, and improving the alleviation of disease states. As used herein, the term "delaying disease progression" means delaying, hindering, slowing, arresting, stabilizing, inhibiting, and/or delaying the progression of a disease. This delay may have a different length of time depending on the disease being treated and/or the history of the individual. As used herein, in the context of administration, the term "effective amount" of an agent (eg, a pharmaceutical formulation, cell, or composition) refers to an amount effective to achieve the desired result at the desired dosage/amount for a desired period of time. As used herein, the term "therapeutically effective amount" of an agent (eg, a pharmaceutical formulation or cell) refers to the effective delivery of a desired therapeutic result (such as for treating a disease, condition, or condition, at a desired dosage for a desired period of time), and / or the amount of pharmacokinetic or pharmacodynamic effect of the treatment). As used herein, "first dose" is used to describe the timing of a given dose prior to administration of a continuous or subsequent dose. The term does not necessarily imply that an individual has never received a dose of cell therapy before or even has never received a single dose of the same cell. As used herein, the term "subsequent dose" refers to a dose administered to the same individual after a previous dose (eg, the first dose) during which no insertion dose is administered to the individual. As used herein, the term "individual" is a mammal, such as a human or other animal, and is typically a human. In some embodiments, the individual has been treated with a therapeutic agent that targets the disease or condition prior to administration. As used herein, the term "pharmaceutical formulation" refers to a formulation that allows for the biological activity of the active ingredient contained therein to be effective, and which does not contain additional components that are unacceptably toxic to the individual administering the formulation. . As used herein, the term "pharmaceutically acceptable carrier" means a component of a pharmaceutical formulation that is non-toxic to the individual other than the active ingredient. Pharmaceutically acceptable carriers include, but are not limited to, buffers, excipients, stabilizers or preservatives. In one aspect, the invention provides a method for treating a polyglutaminic acid (polyQ) disease in a subject, the method comprising administering to the individual parentally or topically an effective amount of stem cells as a unit dose, wherein The administration is carried out in one or more treatment cycles, one of which comprises administering three unit doses at a dosing interval of 2 to 6 weeks, respectively. In some embodiments, polyQ diseases include, but are not limited to, spinocerebellar ataxia (SCA); Machado-Joseph disease (MJD/SCA3); Huntington's disease (HD); dentate red nucleus pallidus Body atrophy (DRPLA); and type 1 X-linked spinal cord myogenic atrophy (SMAX1/SBMA). In some embodiments, the SCA is polyglutamate (polyQ) mediated SCA, and the SCA is preferably SCA1, SCA2, SCA3, SCA6, SCA7 or SCA17. More preferably, the SCA is SCA3. In some embodiments, the mesenchymal stem cell line is a mesenchymal stem cell population (MSC), an adipose tissue derived stem cell (ADMSC) population, an orbital adipose-derived stem cell (OFSC) population, or a four-potential positive stromal cell (QPSC) population. In one embodiment, QPSC is described in US Patent Application No. 14/615,737, QPSC, which has at least 70% cellular homology and exhibits cellular markers CD273, CD46, CD55, and CXCR4, but does not exhibit CD45; Stronger than 70% strength. In one embodiment, the ADSCs are described in US OF 20120288480, which exhibit at least CD90, CD 105, CD29, CD44, CD49b, CD49e, CD58, and HLA-ABC, but not CD133, CD31, CD106, CD146 , CD45, CD14, CD117. The stem cells are preferably a QPSC population. In some embodiments, the cells can be administered by parenteral administration or topical treatment (such as intracerebral or intracranial administration). Parenteral infusions include intramuscular, intravenous, intraarterial or subcutaneous administration. Parenteral administration is preferably intravenous. In some embodiments, the unit dose in 0.5 × 10 5 to 5 × 10 10 cells / kg of body weight. In some embodiments, the unit dose is from 0.5 x 10 5 to 5 x 10 9 , from 0.5 x 10 5 to 5 x 10 8 , from 0.5 x 10 5 to 5 x 10 7 , from 0.5 x 10 5 to 5 x 10 6 , 1.0. ×10 5 to 5 × 10 10 , 1.0 × 10 5 to 5 × 10 9 , 1.0 × 10 5 to 5 × 10 8 , 1.0 × 10 5 to 5 × 10 7 or 1.0 × 10 5 to 5 × 10 6 cells /kg body weight range. In one embodiment, the administration is performed in one or more treatment cycles, wherein one treatment cycle comprises 2 to 6 weeks, respectively (ie, two weeks, three weeks, four weeks, five weeks, or six weeks. In another In the examples, the dosing interval of two weeks was given three unit doses. The number of treatment cycles of the present invention is determined by the scale for the assessment and rating of ataxia (SARA) (Subramony SH., SARA--a new clinical scale for the assessment and rating of ataxia. Nat Clin Pract Neurol. 2007; 3(3): 136-7; Kim BR, Lim JH, Lee S, Park S, Koh SE, Lee IS, Jung H, Lee J. Usefulness of the Scale for the Assessment and Rating of Ataxia (SARA) in Ataxic Stroke Patients. Ann Rehabil Med . 2011; 35: 772-780; and Tan S, Niu HX, Zhao L et al., Reliability and validity of the Chinese version of the Scale for Assessment and Rating of Ataxia. Chin Med J. 2013; 126(11): 2045-8). SARA is a clinical scale based on semi-quantitative assessment of the level of damage in the cerebellar ataxia (spinal cerebellum, Friedreich, and sporadic ataxia). SARA is based on a scale of performance of 8 items, with a total score of 0 (no ataxia) to 40 (most severe ataxia). These scores are based on gait, span, sitting, speech disorder, finger chase, nose-finger test, rapid alternating hand movement, and sputum-sliding patient performance. After the first treatment cycle, if the individual maintains a SARA total score greater than 5 minutes a month, a second and subsequent treatment cycle will be performed. A unit dose of stem cell line is administered at intervals of 2 to 6 weeks. A 2 to 6 week interval means that the unit dose of the stem cell line is administered once every two weeks, three weeks, four weeks, five weeks, or six weeks. In one embodiment, the dosing interval is once every two weeks. In one embodiment, administration once every two weeks means that the unit dose of stem cell line is administered once every two weeks, i.e., once during the 14 day period, preferably on the same day every two weeks. In a bi-weekly dosing regimen, unit doses are administered approximately every 14 days. In the case of stem cell therapy, administration of a unit dose comprises administering a given amount or number of cells in a single composition and/or a single uninterrupted administration (e.g., a single injection or continuous infusion). In some embodiments, the cell line is part of a combination therapy, such as concurrently with another therapeutic intervention or sequentially in any order. In some embodiments, the stem cell line is co-administered simultaneously with one or more additional therapeutic agents or in combination with another therapeutic intervention, or sequentially in any order. In some cases, the additional therapeutic agent or another therapy is co-administered with the cells in close enough time to thereby effect the additional therapeutic agent or another therapy to enhance the effect of the population of cells, or vice versa. In some embodiments, the stem cell line is administered prior to the one or more additional therapeutic agents. In some embodiments, the stem cell line is administered after the one or more additional therapeutic agents. The stem cell line for use in the methods of the invention is formulated as a pharmaceutical composition or formulation, such as a unit dosage form composition comprising the number of cells for administration of a given dose or portion thereof. Pharmaceutical compositions and formulations generally include one or more optional pharmaceutically acceptable carriers or excipients. In some embodiments, the composition includes at least one additional therapeutic agent. The selection of the carrier is determined in part by the particular stem cells and/or by the method of administration. Therefore, there are a variety of suitable formulations. For example, the pharmaceutical composition can contain a preservative. The pharmaceutically acceptable carrier is generally non-toxic to the recipient at the dosages and concentrations employed, and includes, but is not limited to, buffers such as phosphates, citrates, and other organic acids; antioxidants, including ascorbic acid and methyl sulfide. Aminic acid; preservative (such as octadecyldimethylbenzylammonium chloride; hexahydroxyquaternium chloride; benzalkonium chloride; benzethonium chloride; phenol, butanol Or benzyl alcohol; alkyl p-hydroxybenzoate, such as methyl or propyl p-hydroxybenzoate; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol; Low molecular weight polypeptide; protein such as serum albumin, gelatin or immunoglobulin; hydrophilic polymer such as polyvinylpyrrolidone; amino acid such as glycine, glutamic acid, aspartame, histamine Acid, arginine or lysine; monosaccharides, disaccharides, and other carbohydrates, including glucose, mannose or dextrin; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; Salt formation relative to ions, such as sodium; metal mismatch (E.g., Zn- protein complexes); and / or non-ionic surfactant, such as polyethylene glycol (PEG). In some aspects, a buffering agent is included in the composition. Suitable buffering agents include, for example, citric acid, sodium citrate, phosphoric acid, potassium phosphate, and various other acids and salts. In some aspects, a mixture of two or more than two buffers is used. Methods of preparing pharmaceutically acceptable pharmaceutical compositions are known. Formulations can include aqueous solutions. The formulation or composition may also contain more than one active ingredient that can be used in a particular indication, disease or condition for treatment with stem cells. In some embodiments, a pharmaceutical composition comprises stem cells in an amount effective to treat a disease or condition, such as a therapeutically effective amount. In some embodiments, therapeutic efficacy is monitored by periodic assessment of the individual being treated. The desired dose can be delivered by administering the cells by a single bolus injection, by administering the cells by multiple bolus injections, or by administering the cells by continuous infusion. Stem cells and compositions can be administered using standard dosing techniques, formulations, and/or devices. The administration of stem cells can be either autologous or heterologous. Sterile injectable solutions can be prepared by incorporating the cells into a solvent, such as mixing the cells with a suitable carrier, diluent or excipient such as sterile water, physiological saline, dextrose, dextrose or the like. preparation. The composition may contain auxiliary substances such as wetting agents, dispersing or emulsifying agents (for example methylcellulose), pH buffering agents, gelling or viscosity enhancing additives, preservatives, flavoring agents, depending on the route of administration and the desired formulation. And / or colorants. In some aspects, reference may be made to standard text to prepare suitable formulations. Various additives may be added to enhance the stability and sterility of the composition, including antimicrobial preservatives, antioxidants, chelating agents, and buffers. Prevention of the action of microorganisms can be ensured by various antibacterial and antifungal agents (for example, parabens, chlorobutanol, phenol and sorbic acid). The absorption of injectable pharmaceutical forms can be extended by the use of absorption delaying agents such as aluminum monostearate and gelatin. EXAMPLES I. ANIMAL MODEL TEST MATERIALS AND METHODS Animals and experimental design MJD84.2 (B6; CBA-Tg(ATXN3*)84.2Cce/IbezJ) mice without significant motor function degradation have been established as human Machado-Joseph Disease, also known as the disease model of type 3 spinal cerebellar ataxia (MJD/SCA3). In the present study, MJD84.2 animals aged between 20 and 34 weeks were studied. Behavioral analysis of these animals, including improved SHIRPA, footprint analysis, and spin test. Three, test subject injections were performed on 21, 23 and 25 week old mice at two week intervals. The research design is outlined below. A total of thirteen SCA3 Tg/0 mice (B6; CBA-Tg (ATXN3*) 84.2Cce/IbezJ) and eight C57BL/60 0/0 wild-type mice were all derived from JAX with significant motor function degradation. laboratory. Animals were randomly enrolled into four experimental groups: (1) SCA3+ cells; (2) SCA3+PBS; (3) Wt+ cells; (4) Wt+PBS. Three test article injections were performed at two week intervals after determining that the SCA3 Tg/0 mice had a significant disease phenotype. The research design is outlined below. Mesenchymal Stem Cells The QPSC in this study is human ADMSC (Stemchymal®), a cellular product manufactured by Steminent Biotherapeutics Inc. (SBI). In vitro culture amplification and quality control of ADMSCs were performed in a cell factory established according to the PIC/S Good Manufactory Practice guideline following SBI standard operating procedures. Briefly, adipose tissue was collected from healthy donors and immediately delivered to the SBI processing facility at low temperatures (0-5 °C). ADMSCs were isolated, purified and maintained in SBI-unique medium during culture expansion. The 12th generation ADMSC is a high-level QPSC with CD273, CD46, CD55 and CXCR4, which is then packaged in a cryopreservation bag, and the product (Stemchymal®) is delivered for quality testing and cryopreservation. Stemchymal® quality control consists of in-process control and product release testing including, but not limited to, viability, sterility, mycobacterial testing, endotoxin assessment, MSC phenotype (for CD 73, CD 90 and CD) 105 positive, negative for CD 34, CD 45, CD11b, CD 19 and HLA-DR) and three-lineage differentiation ability (osteogenic differentiation, chondrogenic differentiation and adipogenic differentiation). For studies using non-cell given significant degeneration of motor function in mice: A 2.5 × 10 7 cells / kg body weight Stemchymal ® thawed prepared and loaded into a 1 ml insulin syringe (29 1 / 2G). Slowly inject the cells within one hour of thawing (duration of 15 to 20 seconds). For studies using mice with significant motor function degradation : mice were randomized into four groups: (1) SCA3+ cells; (2) SCA3+ PBS; (3) Wt+ cells; (4) Wt+PBS. The inner Stemchymal 2.5 × 10 7 cells / kg body weight of each mouse ® intravenous infusion of 3 to the first group and second group. A total of 125 μl of cell suspension (1:1 low temperature solution (Biolife)) or PBS (Gibco) was administered to each mouse. For both studies, animals were monitored for 4 hours after injection and observed daily. Cells were administered once every two weeks for a total of three times. Data collection and analysis Mice were sacrificed one month after the last test article injection. The body weight of the mice and the latency to fall were recorded throughout the study. Mouse footprints were also analyzed for gait performance after test article injection. Mouse tissues (cortex, cerebellum, heart, kidney, liver, spleen, lung, and tail) were collected for future histopathological analysis and biological distribution studies. Statistical data were presented as mean ± SEM. The results of the spin and foot test were analyzed using the Student's t-test and the significance threshold of p < 0.05. Motion coordination and balance analysis Motion coordination and balance were evaluated in a rotating rod device (MK-670, Muromachi Kikai Co., Ltd., Japan). The mice were placed on a rotating rod at a constant speed (4 rpm) which was accelerated to 40 rpm over a 5 min period. Record the wait time for the drop or wait for the complete passive rotation (attach to the full rotation on the rod). Mice were tested 3 times with 15 min breaks between trials. The average waiting time of each mouse in each test was calculated. The test results were statistically analyzed using the t test. SHIRPA Test The modified SHIRPA test was performed at 20, 24, and 28 weeks of age in mice. The SHIRPA program was adapted from the improved SHIRA program of RIKEN BRC. Test items and scoring criteria are listed in the table below. The mice were scored according to their behavior. The total number of mice in the study group was calculated as 100%. Results are presented as a percentage of the number of mice at certain scoring levels. Footprint analysis Mouse footprints were analyzed approximately one month after the last administration of the cells. Referring to the paper published in 2015 (1), the mouse foot is immersed in the ink (forefoot: red; hind foot: green) so that the mouse leaves a footprint when walking along the corridor or running to the target box. The mice will be placed on a piece of paper (50 cm long, 10 cm wide) in front of the tunnel. Measuring stride, sway, span length, and front and rear paws overlap to indicate gait (see figure below). All mice were subjected to three rounds of measurement before sacrifice. MTT analysis of CD3 + T cell isolation Human peripheral blood mononuclear cells (PBMC) were isolated from heparinized whole blood from healthy donors by Histopaque-1077 (Sigma-Aldrich) density gradient centrifugation. Subsequently, CD3 + T lymphocytes were purified from PBMC by positive selection using magnetic particles (BD Biosciences) conjugated with anti-human CD3 antibody following the manufacturer's instructions. T cell proliferation assay Purified human CD3 + was stimulated in a 96-well plate using disc-conjugated anti-CD3 monoclonal antibody (2 μg/ml) and anti-CD28 monoclonal antibody (2 μg/ml) (BD Biosciences) . T cells (1 × 10 5 cells) and co-cultured with different amounts of ADMSC in 10% fetal bovine serum (FBS), 2 mM l-glutamic acid, 100 U/ml penicillin, 100 U/ml Streptomycin and 25 mM HEPES in RPMI-1640 medium (Gibco). After 48 hours, 5-bromo-2-deoxyuridine (BrdU) was added to each well and the plate was incubated for an additional 18 hours to measure T cell proliferation. The amount of BrdU incorporated into the T cells was measured using a Cell Proliferation ELISA BrdU kit (Roche) according to the manufacturer's instructions. Immunohistochemistry ( IHC ) To evaluate the neuroprotective effects of QPSC, QPSC was injected via the tail vein of the C57BL/6J SCA2 transgenic mouse (IV hMSC-Tg group) or via the occipital foramen (IC hMSC-Tg group). Cerebellar position. A specific antibody (Abcam, code: ab15976) that reacts with human β2 microglobulin is selected to display human cells in murine brain tissue by IHC. Mouse slices (4 μm) were cut and placed on a microscope slide. The sections were rehydrated by rinsing twice in xylene, 100% ethanol, 95% ethanol, and 80% ethanol at 5 min intervals. After paraffin removal, sections were treated with 3% H 2 O 2 to inactivate peroxidase, heated in 10 mM citrate buffer (containing 0.05% Tween 20) for antigen retrieval, and blocked with 1% Solution blocking (1% BSA and 0.1% Triton X-100 (in PBS), Chang et al, Journal of Biomedical Science 2011, 18:54 http://www.jbiomedsci.com/content/18/1/54 , page 3 of 9). The sections were incubated with specific anti-human b2 microglobulin polyclonal antibody (Abeam) diluted in blocking solution (1:400) for 40 min at room temperature. After washing thoroughly with PBS three times, the sections were incubated with secondary antibodies diluted (1:1000) in blocking solution for 40 min at room temperature. Primary antibodies were detected using the EnVision Detection System (DAKO) and observed with diaminobenzidine (DAB; DAKO). The staining was contrasted with aqueous hematoxylin (Sigma Aldrich). For direct comparison, all slides were processed in batches to minimize variability. Safety Test Animals C57BL/6 mice received 3 doses of QPSC via a tail vein injection using an insulin syringe 1/2 cc 30G x 3/8" needle (Terumo, or BD Bioscience). Prior to injection, the animals were warmed for 15 to 20 minutes by a heating pad placed under the cage to dilate their tail veins. Prior to autopsy, all animals were anesthetized with urethane (2 g/kg body weight, Sigma-Aldrich) followed by blood collection from the submandibular vein or via cardiac puncture. Blood sample collection For hematology analysis, whole blood samples were collected in EDTA-containing blood collection tubes (BD Bioscience, Cat. No. 365974). For blood chemistry analysis, whole blood samples were collected in a blood collection tube (BD Bioscience, Cat. No. 365967) containing a plasma separator. After standing the plasma tube for 20 min at room temperature, the plasma was then separated by centrifugation at 6000 rpm for 5 min at 4 °C. Visual autopsy and tissue collection After the blood sample is taken, the animal organs are collected. They were divided into two parts: (1) half of the organ was stored in a -80 ° C freezer, then transferred and stored in a liquid nitrogen container for biodistribution analysis; (2) the other half was fixed (4% paraformaldehyde) , Sigma-Aldrich) and paraffin embedded for histopathological analysis. Quantitative PCR Following the manufacturer's instructions, total RNA was extracted from QPSC or murine tissue using a Total RNA Miniprep Purification Kit (GMbiolab Cat. No. TR01). Subsequently, cDNA synthesis was performed using a two-step MMLV RT-PCR kit (GMbiolab Cat. No. RP012-M). Quantitative PCR was performed using The Fast SYBR® Green Master Mix (Thermo Cat # 4385612) to analyze the relative performance of the selected genes. ELISA was used to measure the intracellular and secreted levels of EGF, FGF-b, VEGF, PDGF and TGF-b1 in QPSC. Samples of cell lysates and modified media were prepared as follows: QPSC was dissolved by freeze-thaw method and at speeding The supernatant of the cell lysate was collected after centrifugation. And for improved media collection, the media was collected after 3 days of QPSC culture. Finally, the concentration of the above growth factors was determined by ELISA (R&D system) according to the manufacturer's instructions. Neuronal cell co-culture test Human astrocyte cell line SVG p12 was treated with 1250 μM 1-methyl-4-phenylpyridine (MPP+) and with different ratios of QPSC (SVG p12: QPSC=1:0.1-1:10) )Co-culture. After 24 hours, the number of cells of SVG p12 was calculated. Example 1 QPSC alters the phenotype of SCA3 QPSC alters the phenotype of SCA3 mice. As shown in Figure 1, SCA3 mice exhibited a slightly broader base compared to wild-type mice and after QPSC treatment, the appearance of SCA3 mice looked similar to wild-type mice. Similar improvements were observed in various functional tests, such as the modified SHIRPA (Fig. 4, Fig. 5), footprints (Fig. 6, Fig. 7), and the spin performance analysis (Fig. 5C). Example 2 The treatment of the present invention stopped weight loss and had no side effects on organ tissues. QPSC also stopped weight loss of SCA3 during disease progression (Fig. 3). Nonetheless, the 3 doses of QPSC did not affect the type of whole blood count (Table 1) or blood biochemistry (Table 2) of SCA3 individuals. Tables 1 and 2 show that the whole blood count/biochemical profile of wild type mice of 25 to 30 weeks old did not differ between the three doses of QPSC and physiological saline (three doses per week). Histopathological analysis revealed normal findings of various vital organ tissues after three doses of QPSC were injected (Figure 2). Table 1 Data are presented as mean ± SD. Table 2 Data are presented as mean ± SD. Example 3 Immunomodulation and anti- ROS ability in mice treated with the present invention and the performance of multiple neurotrophic factors and growth factors. In vitro studies have shown that QPSC not only has immunomodulatory and anti-ROS capabilities ( Figure 10), and also has the ability to express multiple neurotrophic factors and growth factors (Figure 12). In vivo studies showed that SCA mice exhibited improved spine performance under oxidative stress after QPSC treatment (Figure 11). In addition, QPSC can also prevent neuronal loss both in vitro (Figure 13) and in vivo (Figure 14). Although questions have been raised regarding the possibility of cell migration across the blood-brain barrier (BBB), the ability of QPSC to be located intracranially via intravenous infusion has been shown (Figures 8 and 9). Therefore, it is reasonable to conclude that via intravenous infusion, QPSC can reach the cerebellum through the BBB and protect neuronal cells of SCA individuals from ROS and immune overreaction. QPSC also secretes multiple neurotrophic and growth factors to maintain the number of neuronal cells, thereby delaying poly-Q diseases (such as polyglutaminic spinocerebellar ataxia, Machado-Joseph disease, Huntington's disease, DRPLA) And the progress of SMAX1/SBMA). II. Human Clinical Trials This human clinical trial aims to study Stemchymal® infusion for the treatment of polyglutamin-mediated diseases (such as polyglutamate-induced spinal cord) by a randomized, double-blind, placebo-controlled study design. Therapeutic efficacy and safety of cerebellar ataxia, Machado-Joseph disease, Huntington's disease, DRPLA and SMAX1/SBMA. Eligible individuals will receive Stemchymal® via intravenous infusion. In one example of spinocerebellar ataxia against polyglutamate, the individual undergoing the trial has genotype-confirmed type 2 spinocerebellar ataxia or type 3 spinocerebellar ataxia. The individual's baseline SARA score is in the range of 5 to 15 minutes. The 2.5 × 10 7 cells / kg body weight Stemchymal ® thawed prepared and loaded into a syringe. Slowly inject the cells within one hour of thawing. The Stemchymal ® by intravenous infusion to each subject and at intervals of every two weeks for the first three administrations cells. After one or more treatment cycles, the individual's SARA score decreases and the SCA2 or SCA3 condition improves.