2012-06-012024-05-17https://scholars.lib.ntu.edu.tw/handle/123456789/681104摘要：Dravet Syndrome是小兒肌張力型癲癇症候群(severe myotonic epilepsy of infant type, SMEI)，病患通常在新生兒時期已經發病，且因對藥物治療有抗藥性故預後極差。文獻顯示此疾病與基因SCN1A所轉錄之第一型鈉離子通道(Nav1.1 channel)突變有關。有鑑於台灣的SMEI病患亟需有效之臨床醫療，本研究將以基因替換小鼠(knock- in mice)做為癲癇治療藥物之篩選平台，期以能對此型癲癇提出有效之治療方式。文獻顯示大部份的Dravet Syndrome是由於原生性的(de novo)在SCN1A此a type I鈉離子通道蛋白上發生突變，目前約有一百多種異型合子突變被發現，包括在台灣也有相關病例，為了尋找更適合的治療策略，我們將針對東方人疾病模型製備基因替換小鼠(E1099X)，成為有潛力的藥物篩選平台。 在癲癇患者身上，常可發現多種離子通道的基因產生突變。例如一種顯性遺傳的全身性癲癇併發熱痙(GEFS+)—表現在幼童為熱痙攣，成人為非熱痙攣—即可發現病患具有神經細胞鈉離子電位感知通道的α-1次單位SCN1A基因，發生錯義突變。SCN1A的重組變異也被認為與新生兒的肌僵直性癲癇(SMEI)有關，SMEI的初徵為發熱，而後非特定型態的全身性或局部痙攣(包括肌肉僵直)，稍後表現心理運動能力的延遲。SMEI是一種罕見疾病，無法以藥物治療，直至今日，已有許多SCNIA突變的幼童被診斷為SMEI。在本計畫中，我們將關注在SCN1A的特定片段—E1099X基因錯義突變上。我們計畫利用基因轉殖小鼠，表現SCN1A的E1099X基因錯位突變，針對concordant antisense oligonucleotides新型藥物對於此SCN1A突變小鼠之治療成效及其藥理作用，包含劑量的選擇、藥物efficacy試驗、增加SCN1A在腦部各區域表現量、行為表現的評估 (behavioral assessments)。 <br> Abstract: Dravet Syndrome (severe myotonic epilepsy of infant, SMEI) is a kind of rare disease in clinic. The patients usually have early onset during neonatal period and poor outcome due to refractory to classical antiepileptic medical therapy. Studies indicate that mutation of scn1a gene, which encodeds Nav1.1 channels in central nervous system, is related with SMEI. In order to find out effective clinical medical therapy for the patients in Taiwan, current project will use Scn1a knock-in mice as a platform to screen the therapeutic efficacy of four novel compounds in treating patients with Dravet syndrome. The onset of the disease typically occurs within the first year, with prolonged, generalized, or unilateral clonic seizures triggered by fever. They are frequently categorized as febrile seizures. Literature search results indicate that most of the patients with Dravet syndrome are resulted from de novo point mutations in a voltage-gated sodium channel type Iα (SCN1A) gene. There are more than 300 different heterozygous SCN1A mutants identified worldwide, including cases from Taiwan. To optimize the therapeutic formulation, we built a drug testing platform using a mutant mouse model carrying an Asian-specific point mutation allele which leads to a premature translation termination (E1099X). SMEI is a rare disorder and is refractory to drug therapy. To date, many kinds of SCN1A mutations have been reported in SMEI patients. In this proposal a specific missense mutation of the SCN1A (E1099X) will be focused. The specific aim of current project is to screen the efficacies of one concordant antisense oligonucleotide on the SCN1A mutation-induced epileptogenesis. This concordant antisense oligonucleotide, CUR-1901, will be provided by Dr. Jane Hsiao (許照惠博士). The pharmacological properties and therapeutic efficacy, including determination of the suitable dosages (determination of the the dose-dependent curve), the effects of increases in the SCN1A expression in several brain structures, and the assessments of animal behaviors before and after treatment, will be elucidated in this study.Dravet syndromeConcordant antisenseTransgenic miceEpilepsyDravet syndromeConcordant antisenseTransgenic miceEpilepsy