Whole blood gene expression to identify biomarkers of disease risk, progression and response to therapy in Type 1 diabetes

摘要 在这项资助中，我们建议对外周血细胞进行基因表达分析 自身抗体阴性的T1D患者一级亲属(AA-FDRs)和AA+的PBC 已经进展(进展者)或没有进展到高血糖(非 进展者)，与健康的非T1D相关对照组比较。我们还将表演基因 新入选TrialNet abatacept(CTLA4-Ig)患者外周血淋巴细胞的表达分析 发病研究(TN-09)。拟议的研究将确认和扩大我们在 儿童疾病易感性和疾病进展的全血生物标志物鉴定 T1D，并启动对治疗反应的生物标志物的研究。 在JDRF的资助下产生的初步数据表明，PBC基因 T1D和T1D相关个体的表达谱彼此更相似，而不是 健康的非糖尿病相关的正常对照；也就是说，有前驱基因表达 可以识别T1D患者AA-FDR的PBCs中疾病风险的签名 疾病的临床征兆。根据这些初步数据，我们假设T1D家族 患者，都有环境驱动的、表观遗传控制的基因表达特征 在外周血液中可以看到的风险。我们确定了AA-FDR的一个子集 干扰素诱导的外周血细胞基因表达水平升高。这些人还 表达了一个更类似于AA+FDR的基因表达特征 与无进展的AA+FDR或其他AA-FDR和对照组相比， 提示全血基因表达生物标记物可用于识别个体 世卫组织将在血清转换前进展为高血糖。在血清转换患者中 (AA+)，我们发现基因表达在疾病的早期阶段变化最大(&gt；3 进展期患者与非进展期患者相比，在高血糖发作前数年)。这 表明我们可能能够识别出不仅区分AA+进展因子的生物标志物 来自AA+的非进展者，还可以预测疾病的进展率。最后，研究 已表明新近发病的高血糖患者的PBC基因表达特征不同 与那些没有反应的人相比，对抗CD3治疗有反应的人(ABATE试验)。我们 假设我们可能也看到患者在对 CTA4-Ig(Abatacept)与那些在开始治疗3个月后没有反应的人进行比较 治疗(TN-09)。如果能够及早发现无反应者，这将为其他人留出时间 在β细胞质量/功能完全丧失之前测试的治疗方法。 提出了三个具体目标。第一个特定的目标是验证前驱症状 糖尿病相关AA患者的基因表达模式。第二个具体目标是 确定T1D疾病进展的PBC基因表达特征(确定 谁将从那些不会进步到T1D的人)，以及进步速度的签名。这个 第三个特定目标是询问是否有PBC基因表达特征可以预测 近期发病中CTLA4-Ig干预的应答者与非应答者的表型比较 高血糖患者使用TN09样本。本申请书中描述的研究需要 来自TrialNet的PBC RNA样本。我们已经收到了来自TrialNet的&gt；300个样品 储存库，并已获得临时批准以接收额外样本(信函 附连)。

Summary In this grant, we propose to perform gene expression analysis on peripheral blood cells (PBC) of auto-antibody negative first degree relatives of T1D patients (AA- FDRs) and AA+ FDRs who have either progressed (progressors) or not progressed to hyperglycemia (non- progressors), compared to healthy non-T1D related controls. We will also perform gene expression analysis on PBCs of patients enrolled in the TrialNet Abatacept (CTLA4-Ig) New Onset Study (TN-09). The proposed studies will confirm and extend our work on the identification of whole blood biomarkers of disease susceptibility and disease progression in T1D, and initiate our studies on biomarkers of response to therapy. Preliminary data generated under funding from the JDRF suggests that the PBC gene expression profiles of T1D and T1D-related individuals are more similar to each other than to healthy normal non-diabetes-related controls; that is, there is a prodromal gene expression signature that can identify disease risk in the PBCs of AA- FDRs of T1D patients who show no clinical signs of disease. Based on these preliminary data, we hypothesize that families of T1D patients, all have an environmentally-driven, epigenetically-controlled gene expression signature of risk that can be seen in peripheral blood. We identified a subset of AA- FDRs that had elevated levels of interferon-induced gene expression in their PBCs. These individuals also expressed a gene expression signature that was more similar to AA+ FDRs who progressed to hyperglycemia than to AA+ FDRs who did not progress, or to other AA- FDRs, and to controls, suggesting that whole blood gene expression biomarkers could be used to identify individuals who will progress to hyperglycemia prior to seroconversion. In patients who seroconverted (AA+), we found that gene expression was most changed during the early stages of disease (>3 years before the onset of hyperglycemia) in progressors compared to non-progressors. This indicates that we might be able to identify biomarkers that not only distinguish AA+ progressors from AA+ non-progressors, but also predict the rate of disease progression. Finally, studies have shown that PBC gene expression signatures differ in recent onset hyperglycemic patients that responded to anti-CD3 therapy compared to those who did not respond (AbATE trial). We hypothesize that we may also see a difference in gene expression in patients who respond to CTA4-Ig (Abatacept) compared to those who do not respond, 3 months after the initiation of therapy (TN-09). If non-responders could be identified early, this would allow time for other therapies to be tested before total loss of beta cell mass/function occurs. Three specific aims are proposed. The first specific aim is to validate the prodromal gene expression pattern seen in diabetes related AA- individuals. The second specific aim is to identify both a PBC gene expression signature of T1D disease progression (identifying those who will progress to T1D from those who will not), and a signature of rate of progression. The third specific aim is to ask if there is a PBC gene expression signature that might predict responder vs. non-responder phenotypes for intervention with CTLA4-Ig in recent onset hyperglycemic patients using TN09 samples. The studies described in this application require PBC RNA samples from TrialNet. We have already received >300 samples from the TrialNet repository and have received provisional approval to receive additional samples (letter attached).