Ocampo-Ortega Sergio Adrian, Sierra-Sanchez Vivany Maydel, Blancas-Napoles Citlali Margarita, González-Carteño Asdrúbal, Mera-Jiménez Elvia, Macías-Pérez Martha Edith, Hernandez-Guerra Adriana, Romero-Nava Rodrigo, Huang Fengyang, Hong Enrique, Villafaña Santiago
Laboratorio de Terapia Génica Experimental, Escuela Superior de Medicina, Instituto Politécnico Nacional, Ciudad de Mexico 11340, Mexico.
Laboratorio de Cultivo Celular, Neurobiología y Conducta, Escuela Superior de Medicina, Instituto Politécnico Nacional, Ciudad de Mexico 11340, Mexico.
Life (Basel). 2024 Dec 4;14(12):1607. doi: 10.3390/life14121607.
Huntington's disease is a genetic disorder characterized by progressive neuronal cell damage in some areas of the brain; symptoms are commonly associated with chorea, rigidity and dystonia. The symptoms in Huntington's Disease are caused by a pathological increase in the number of Cytokine-Adenine-Guanine (CAG) repeats on the first exon of the Huntingtin gene, which causes a protein to have an excessive number of glutamine residues; this alteration leads to a change in the protein's conformation and function. Therefore, the purpose of this work was to design, synthesize and evaluate an antisense oligonucleotide (ASO; 95 nucleotides) HTT 90-5 directed to the Huntingtin CAG repeats in primary leukocyte culture cells from a patient with Huntington's Disease; approximately 500,000 leukocytes per well extracted from venous blood were used, to which 100 pMol of ASO were administered, and the expression of Huntingtin was subsequently evaluated at 72 h by RT-PCR. Our results showed that the administration of the HTT 90-5 antisense decreased the expression of Huntingtin mRNA in the primary culture leukocyte cells from our patient. These results suggest that the use of long antisense targeting the CAG Huntingtin cluster may be an option to decrease the expression of Huntingtin and probably could be adjusted depending on the number of CAG repeats in the cluster.
亨廷顿舞蹈症是一种遗传性疾病,其特征是大脑某些区域的神经元细胞进行性损伤;症状通常与舞蹈症、僵硬和肌张力障碍有关。亨廷顿舞蹈症的症状是由亨廷顿基因第一个外显子上细胞因子 - 腺嘌呤 - 鸟嘌呤(CAG)重复序列数量的病理性增加引起的,这导致一种蛋白质含有过多的谷氨酰胺残基;这种改变会导致蛋白质的构象和功能发生变化。因此,本研究的目的是设计、合成并评估一种针对亨廷顿舞蹈症患者原代白细胞培养细胞中亨廷顿CAG重复序列的反义寡核苷酸(ASO;95个核苷酸)HTT 90 - 5;每孔使用从静脉血中提取的约500,000个白细胞,并向其中加入100 pMol的ASO,随后在72小时通过逆转录聚合酶链反应(RT - PCR)评估亨廷顿蛋白的表达。我们的结果表明,HTT 90 - 5反义寡核苷酸的给药降低了我们患者原代培养白细胞中亨廷顿蛋白mRNA的表达。这些结果表明,使用靶向CAG亨廷顿簇的长反义寡核苷酸可能是降低亨廷顿蛋白表达的一种选择,并且可能可以根据簇中CAG重复序列的数量进行调整。