Prevent IgM MGUS Progression by Targeting the Driver Mutation

通过靶向驱动突变预防IgM MGUS进展 项目摘要 IgM类别意义不明的单克隆丙种球蛋白病（IgM MGUS）患者在 Waldenstrom巨球蛋白血症（WM）、非霍奇金淋巴瘤（NHL）、慢性淋巴细胞性 白血病（CLL）或原发性轻链（AL）淀粉样变性。髓样分化因子88（MYD 88）是 在20世纪90年代作为骨髓前体中的初级分化反应基因被发现。错义 MYD 88中265位亮氨酸变为脯氨酸的突变（L265 P）在约90%的WM中发现。此驱动程序 突变也发生在>50%的原发性淋巴结淋巴瘤和~29%的活化B细胞弥漫性大B- 细胞淋巴瘤（ABC-DLBCL），以及约52%的IgM MGUS。除IgM外，无癌前病变-癌对 MGUS-WM具有单个氨基酸取代，如MYD 88 L265 P，其发生在大多数癌前病变中，并且约90%的MGUS-WM具有类似于MYD 88 L265 P的氨基酸取代。 MYD 88 L265 P是研究癌症预防中单一致病突变的典范 和拦截。我们发现RING指蛋白138（RNF 138）与赖氨酸63（K63）连接， 在MYD 88 L265 P上的多聚泛素链，但令人惊讶的是，它对WT MYD 88几乎没有活性。这种翻译后 MYD 88 L265 P上的修饰对其致癌作用至关重要。我们使用了一种深度学习的人工智能， 基于神经网络的人工智能（AI）技术，可以虚拟筛选约1000万种化合物。我们 鉴定了靶向MYD 88中L265 P附近结合位点的主要命中化合物的分数。我们验证 来自AI筛选的主要命中物，并评估它们对MYD 88 L265 P泛素化和异种移植的抑制 肿瘤发生一种化合物用MYD 88 L265 P减弱NHL细胞的淋巴瘤生长，但用MYD 88 L265 P不能。 WT MYD88 E3连接酶RNF 138主要在睾丸和免疫细胞中表达，其整个- 在小鼠中的体基因敲除不影响除睾丸中的生理功能之外的生理功能。RNF 138缺失 减弱来自具有MYD 88 L265 P的WM和DLBCL细胞的肿瘤发生，但不减弱来自具有WT MYD 88的细胞的肿瘤发生。 我们假设靶向MYD 88 L265 P-RNF 138相互作用阻止IgM MGUS进展。在这 应用，我们提出了两个具体的目标，以确定和验证化学和免疫预防剂， 靶向MYD 88 L265 P-RNF 138并防止IgM MGUS进展为WM和NHL。在目标1中，我们将使用 AI开发的MYD 88 L265 P靶向化合物可预防IgM MGUS进展。在目标2中，我们将使用 针对RNF 138的DNA疫苗以防止IgM MGUS进展。我们希望能产生有效的化学物质 以及用于癌症预防和针对IgM MGUS的拦截的无明显毒性的免疫剂 进展

Prevent IgM MGUS Progression by Targeting the Driver Mutation Project Summary Patients with monoclonal gammopathy of undetermined significance of the IgM class (IgM MGUS) are at increased risk for Waldenstrom macroglobulinemia (WM), non-Hodgkin lymphoma (NHL), chronic lymphocytic leukemia (CLL), or primary light-chain (AL) amyloidosis. Myeloid differentiation factor 88 (MYD88) was discovered in the 1990s as a primary differentiation response gene in myeloid precursors. A missense mutation (L265P) changing leucine at position 265 to proline in MYD88 is found in ~90% of WM. This driver mutation also occurs in >50% of primary extranodal lymphomas and ~29% of activated B-cell diffuse large B- cell lymphomas (ABC-DLBCL), as well as in ~52% of IgM MGUS. No precancer-cancer pairs other than IgM MGUS-WM have a single amino acid substitution like MYD88 L265P occurring in most precancer and ~90% of cancer, making MYD88 L265P paradigmatic for the study of a single causative mutation in cancer prevention and interception. We found that the RING finger protein 138 (RNF138) attaches lysine 63 (K63)-linked polyubiquitin chains to MYD88 L265P, yet strikingly, it has little activity on WT MYD88. This posttranslational modification on MYD88 L265P is essential to its oncogenic action. We have used a deep learning artificial intelligence (AI) technology based on a neural network to virtually screen about ten million compounds. We identified scores of primary hit compounds targeting a binding site near L265P in MYD88. We validated primary hits from AI screening and evaluated their inhibition of MYD88 L265P ubiquitination and xenograft tumorigenesis. One compound attenuated lymphoma growth from NHL cells with MYD88 L265P but not that with WT MYD88. The E3 ligase RNF138 is primarily expressed in the testis and in immune cells, and its whole- body knockout in mice does not affect physiological functions other than that in the testis. RNF138 deletion attenuates tumorigenesis from WM and DLBCL cells with MYD88 L265P but not from cells with WT MYD88. We hypothesize that targeting the MYD88 L265P-RNF138 interaction prevents IgM MGUS progression. In this application, we propose two specific aims to identify and validate chemo- and immune-prevention agents to target MYD88 L265P-RNF138 and prevent IgM MGUS progression into WM and NHL. In Aim 1, we will use the AI-developed MYD88 L265P-targeting compound to prevent IgM MGUS progression. In Aim 2, we will use DNA vaccines against RNF138 to prevent IgM MGUS progression. We expect to generate effective chemical and immunological agents without overt toxicities for cancer prevention and interception against IgM MGUS progression.