Forecasting most deleterious nsSNPs in human <i>TLR9</i> gene and their cumulative impact on biophysical features of the protein using <i>in silico</i> approaches

In women, the uterine cervix and corpus uteri are two main suspects, playing a major role in cancer-associated-mortality. Immunologically, Toll-like receptors (TLRs) associated with the innate immune system, can recognize pathogens and induce immune responses against pathogens. Cellularly, TLR9 expression occurs in immune system cells including macrophages, natural killer cells, dendritic cells, and other antigen-presenting cells. TLR9 recognizes and interacts with viral and bacterial DNA comprising cytosine-phosphate-guanine (CpG) dideoxynucleotide motif. The current study is designed to identify the most deleterious nonsynonymous single nucleotide polymorphisms (nsSNPs) in the <i>TLR9</i> gene and to delineate their deleterious effect on the structural and functional features of proteins at the molecular level. Based on the implementation of various computational tools and algorithms eight most deleterious nsSNPs (P139H, R257C, C265Y, L283P, G514D, L544Q, H566Y, and W670R) have been identified in the human <i>TLR9</i> gene as potentially damaging SNPs. Further, our study suggests highly conserved patterns at deleterious nsSNPs sites could influence protein stability and its functional features. Additionally, this study identifies two nsSNPs (G514D and W670R) associated with the severity of Uterine corpus endometrial carcinoma. In support of our computational findings, the validation of key results using polymerase chain reaction and other experimental methods is warranted in the Indian population. In general, this study might be able to delineate the guideline for identifying the most damaging SNPs and enhances the understating of the risk factors for cancer and disease susceptibilities.在女性群体中，子宫颈与子宫体是两大主要的癌变易感部位，其相关癌症在癌症相关死亡中占据主导地位。从免疫学视角来看，与先天免疫系统相关的Toll样受体（Toll-like receptors）可识别病原体，并诱导针对病原体的免疫应答。在细胞层面，Toll样受体9（TLR9）的表达见于巨噬细胞、自然杀伤细胞、树突状细胞及其他抗原呈递细胞等免疫系统细胞中。TLR9可识别并结合含有胞嘧啶-磷酸-鸟嘌呤（cytosine-phosphate-guanine, CpG）二脱氧核苷酸基序的病毒与细菌DNA。本研究旨在鉴定*TLR9*基因中危害性最强的非同义单核苷酸多态性（nonsynonymous single nucleotide polymorphisms, nsSNPs），并在分子水平上阐明这些突变对蛋白质结构与功能特征的有害影响。通过运用多种计算工具与算法，本研究在人类*TLR9*基因中鉴定出8个具有潜在破坏性的nsSNPs：P139H、R257C、C265Y、L283P、G514D、L544Q、H566Y及W670R。进一步研究表明，有害nsSNPs位点处的高度保守模式可能影响蛋白质稳定性及其功能特性。此外，本研究还鉴定出2个nsSNPs（G514D与W670R）与子宫体子宫内膜癌的严重程度相关。为验证本研究的计算结果，在印度人群中采用聚合酶链式反应（polymerase chain reaction）及其他实验方法对关键结果进行验证具有必要性。总体而言，本研究可为鉴定危害性最强的单核苷酸多态性提供指导框架，并加深人们对癌症风险因素及疾病易感性的理解。