Targeting alpha-synuclein with a microRNA-embedded silencing vector in the rat substantia nigra: positive and negative effects.
Targeting alpha-synuclein with a microRNA-embedded silencing vector in the rat substantia nigra: positive and negative effects.
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DOI：
10.1016/j.brainres.2014.01.010
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发表时间：
2014-03-06
期刊：
影响因子：
2.9
通讯作者：
Bohn, Martha C.
中科院分区：
文献类型：
--
作者：
Khodr, Christina E.;Becerra, Amanda;Han, Ye;Bohn, Martha C.
α-突触核蛋白（SNCA）下调显示出对突触核蛋白病（包括帕金森病（PD））的治疗潜力。先前我们表明，在大鼠黑质（SN）中使用短发夹（sh）RNA的人（h）SNCA基因沉默可防止hSNCA诱导的前肢缺陷，但不能防止多巴胺（DA）神经元丢失。此外，miR-嵌入的hSNCA基因沉默shRNA增加了体外细胞死亡，但嵌入microRNA 30转录物（mir 30-hSNCA）中的相同靶序列不会。使用mir 30-hSNCA在体内检查hSNCA基因沉默。用腺相关病毒血清型2/8（AAV）-hSNCA、AAV-hSNCA加AAV-mir 30-SNCA或AAV-hSNCA加对照非沉默mir 30嵌入的siRNA和DA神经元标记物立体定位注射大鼠到一个SN中，并检查相关行为。AAV 2/8介导的SN hSNCA表达诱导前肢缺陷和酪氨酸羟化酶免疫反应性（TH-IR）神经元损失。使用mir 30-hSNCA的hSNCA基因沉默在2个月时防止这种前肢缺陷并改善TH-IR神经元损失。通过蛋白质印迹评估的纹状体（ST）TH-IR纤维密度和DA标志物不受单独的AAV-hSNCA的影响。任一沉默载体的共表达减少了ST TH-IR纤维、SN中的panTH和SN和ST中的Ser 40磷酸化TH，但不影响囊泡单胺转运蛋白-2。然而，hSNCA基因沉默促进部分TH-IR纤维恢复2个月。任一沉默载体的共表达也诱导SN炎症，尽管在hSNCA沉默的SN中通过2 m观察到一些恢复。使用AAV-mir 30-hSNCA的hSNCA基因沉默对前肢行为和SN DA神经元具有积极作用，其受到炎症和减少的TH表达的损害，这表明AAV 2/8-mir 30-hSNCA介导的基因沉默虽然在体外有希望，但不是用于PD的治疗性翻译的候选物。
Alpha-synuclein (SNCA) downregulation shows therapeutic potential for synucleinopathies, including Parkinson’s disease (PD). Previously we showed that human (h)SNCA gene silencing using a short hairpin (sh)RNA in rat substantia nigra (SN) protects against a hSNCA-induced forelimb deficit, but not dopamine (DA) neuron loss. Further, the mir-embedded hSNCA gene silencing shRNA increases cell death in vitro, but the same target sequence embedded in a microRNA30 transcript (mir30-hSNCA) does not. Examine hSNCA gene silencing using mir30-hSNCA in vivo. Rats were stereotaxically injected into one SN with adeno-associated virus serotype 2/8 (AAV)-hSNCA, AAV-hSNCA plus AAV-mir30-SNCA or AAV-hSNCA plus a control non-silencing mir30-embedded siRNA and DA neuron markers and associated behavior were examined. AAV2/8-mediated SN hSNCA expression induces a forelimb deficit and tyrosine hydroxylase-immunoreactive (TH-IR) neuron loss. hSNCA gene silencing using mir30-hSNCA protects against this forelimb deficit at 2m and ameliorates TH-IR neuron loss. Striatal (ST) TH-IR fiber density and DA markers, assessed by western blot, are unaffected by AAV-hSNCA alone. Co-expression of either silencing vector reduces ST TH-IR fibers, panTH in SN and Ser40 phosphorylated TH in SN and ST, but does not affect vesicular monoamine transporter-2. However, hSNCA gene silencing promotes partial TH-IR fiber recovery by 2m. Co-expression of either silencing vector also induces SN inflammation, although some recovery was observed by 2m in hSNCA-silenced SN. hSNCA gene silencing with AAV-mir30-hSNCA has positive effects on forelimb behavior and SN DA neurons, which are compromised by inflammation and reduced TH expression, suggesting that AAV2/8-mir30-hSNCA-mediated gene silencing, although promising in vitro, is not a candidate for therapeutic translation for PD.
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影响因子：
14.9
作者：
Sibley CR;Seow Y;Curtis H;Weinberg MS;Wood MJ
通讯作者：
Wood MJ
DOI：
10.1073/pnas.0711053105
发表时间：
2008-01-15
影响因子：
11.1
作者：
Gorbatyuk, Oleg S.;Li, Shoudong;Muzyczka, Nicholas
通讯作者：
Muzyczka, Nicholas
影响因子：
64.8
作者：
Grimm, Dirk;Streetz, Konrad L.;Kay, Mark A.
通讯作者：
Kay, Mark A.
影响因子：
5.3
作者：
Sapru, MK;Yates, JW;Bohn, MC
通讯作者：
Bohn, MC
影响因子：
15.1
作者：
Lewis, Jada;Melrose, Heather;Bumcrot, David;Hope, Andrew;Zehr, Cynthia;Lincoln, Sarah;Braithwaite, Adam;He, Zhen;Ogholikhan, Sina;Hinkle, Kelly;Kent, Caroline;Toudjarska, Ivanka;Charisse, Klaus;Braich, Ravi;Pandey, Rajendra K.;Heckman, Michael;Maraganore, Demetrius M.;Crook, Julia;Farrer, Matthew J.
通讯作者：
Farrer, Matthew J.