Quantifying proteins in plasma do democratize personalized medicine for patients with type 1 diabetes

量化血浆中的蛋白质确实使 1 型糖尿病患者的个性化医疗民主化

• 批准号: 10730284
• 负责人: ANDREW N HOOFNAGLE
• 金额: $ 86.77万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-21 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10730284
• 关键词: Address; Adoption; Affect; Affinity; Age; Algorithms; Amino Acids; Amputation; Antibodies; Area; Autoimmune Diseases; Autoimmunity; Beta Cell; Biological Assay; Biological Markers; Blindness; Blood Glucose; Blood Tests; Calibration; Characteristics; Child; Chromatography; Chromogranins; Clinical; Clinical Chemistry; Clinical Investigator; Clinical Research; Collaborations; Community Healthcare; Data; Democracy; Deposition; Detection; Development; Diabetes Mellitus; Diagnosis; Digestion; Disease; Emotional; Ensure; Enzyme-Linked Immunosorbent Assay; Epidemiology; Family; Financial Hardship; Genetic; Glucagon; Guidelines; Healthcare Systems; High Pressure Liquid Chromatography; Human; Hybridomas; Hyperglycemia; Hypoglycemia; Immunoassay; Immunologic Factors; In Vitro; Incidence; Individual; Injectable; Institution; Insulin; Insulin-Dependent Diabetes Mellitus; Intervention; Iowa; Islets of Langerhans; Kidney Diseases; Laboratories; Laboratory Study; Lead; Liquid Chromatography; Mass Spectrum Analysis; Measurement; Measures; Methods; Molecular; Monoclonal Antibodies; Myocardial Infarction; National Institute of Diabetes and Digestive and Kidney Diseases; Organ; Pancreas; Patient Care; Patients; Peptides; Periodicals; Persons; Phase; Plasma; Plasmids; Population; Post-Translational Protein Processing; Procedures; Process; Proinsulin; Protein Biosynthesis; Proteins; Proteolysis; Publishing; Reagent; Reproducibility; Research; Research Design; Research Personnel; Risk; Sampling; Serum; Solid; Source; Specificity; Study models; Technology; Testing; Thinness; Translating; Translational Research; Translations; Trypsin; United States; United States National Institutes of Health; Universities; Validation; Western Blotting; Work; analog; biological research; burden of illness; caucasian American; clinical care; detection limit; disorder prevention; disorder risk; experience; experimental study; extracellular vesicles; glycation; improved; instrument; interest; islet amyloid polypeptide; macrovascular disease; method development; mouse model; multiplex assay; novel; personalized medicine; preclinical study; prevent; proglucagon; protein expression; response; stressor; tandem mass spectrometry; tool; validation studies

ABSTRACT Type 1 diabetes affects more than 1.25 million people in the United States and the annual incidence is increasing at an alarming rate of 3-4%. The emotional and financial burden of the disease is overwhelming and we currently have no way to predict or prevent new cases. As we gain a better understanding of the pathophysiological processes in the pancreas and the downstream effects of hyperglycemia (and periodic hypoglycemia during treatment), more robust biomarker assays are needed to improve the reproducibility of research findings and to translate those findings to clinical care. One technology that can provide robust, transferable assays for the measurement of proteins is liquid chromatography-tandem mass spectrometry. By directly detecting the analyte of interest, assays that use mass spectrometry detection can have better specificity than immunoassays and when paired with enrichment strategies, they can also be very sensitive. As we have demonstrated previously, it is straightforward to harmonize the results of mass spectrometric assays, which is significantly more difficult for immunoassays in general. This proposal aims to generate and validate novel transferable protein assays that harness the power of mass spectrometry. We aim to leverage a new method for the enrichment of extracellular vesicles and new de novo proteins for affinity enrichment, called minibinders, to help with sensitivity of the methods. Whenever possible, assays will be multiplexed and if antibodies are required for enrichment, they will be widely distributed through the Iowa Hybridoma Bank. Plasmids encoding minibinders will be deposited at Addgene. Chromatographic data from method development (particularly peptide selection, which will use narrow-window data-independent acquisition rather than relying on algorithms or data- dependent acquisition methods) as well as chromatographic data from method validation will be distributed via Panorama, along with detailed standard operating procedures. As requested in RFA DK-21-031, a portion of the assays produced will target glucagon, other fragments of proglucagon, proinsulin and its fragments, glycated soluble CD59, amylin, and the chromogranins. Our Target Prioritization Committee will help identify the most important proteins to add to this list and focus our development efforts.

抽象的 在美国，1 型糖尿病影响着超过 125 万人，每年 发病率正以 3-4% 的惊人速度增加。的情感和经济负担 疾病势不可挡，我们目前无法预测或预防新病例。正如我们 更好地了解胰腺和胰腺的病理生理过程 高血糖（以及治疗期间周期性低血糖）的下游影响，更多 需要稳健的生物标志物测定来提高研究结果的可重复性并 将这些发现转化为临床护理。一项可以提供稳健、可转移的技术 测量蛋白质的测定方法是液相色谱-串联质谱法。 通过直接检测感兴趣的分析物，使用质谱检测的测定可以 比免疫测定具有更好的特异性，并且当与富集策略结合使用时，它们 也可能非常敏感。正如我们之前所演示的，很容易 协调质谱分析的结果，这对于 一般免疫测定。该提案旨在生成并验证新颖的可转让 利用质谱法进行蛋白质测定。我们的目标是利用一种新方法 用于富集细胞外囊泡和新的从头蛋白质以进行亲和力富集， 称为迷你绑定程序，以帮助提高方法的敏感性。只要有可能，将进行化验 多重使用，如果需要抗体进行富集，它们将通过 爱荷华杂交瘤银行。编码小结合剂的质粒将保藏在 Addgene。 来自方法开发的色谱数据（特别是肽选择，将使用 窄窗口数据独立采集，而不是依赖算法或数据 相关的采集方法）以及方法验证的色谱数据将 通过 Panorama 分发，并附有详细的标准操作程序。按照要求 RFA DK-21-031，所产生的部分检测将针对胰高血糖素、其他片段 胰高血糖素原、胰岛素原及其片段、糖化可溶性 CD59、胰淀素和 嗜铬粒蛋白。我们的目标优先顺序委员会将帮助确定最重要的目标 将蛋白质添加到此列表中并集中我们的开发工作。