Supplement: Bio CaRGOS: Capture and release gels for optimized storage of Cancer Biospecimens

补充：Bio CaRGOS：捕获和释放凝胶以优化癌症生物样本的储存

基本信息

批准号：
10381281
负责人：
Gautam Gupta
金额：
$ 3.5万
依托单位：
UNIVERSITY OF LOUISVILLE
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-01 至 2022-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10381281
关键词：
Achievement Albumins Amylases Antigens Area Biological Markers Buffers Cancer Model Cell Line Chemical Engineering Clinical Complex Cryopreservation Detection Diagnosis Disease Doctor of Philosophy Due Process Early Diagnosis Electrolytes Engineering Equipment Experimental Designs Fibrinogen Formulation Freezing Future Gel Globulins Glucose Goals Grant Immunoassay Individual Ionic Strengths Malignant Neoplasms Malignant neoplasm of pancreas Measures Methanol Methodology Methods Modeling Monitor Nature Patient Monitoring Performance Pharmaceutical Preparations Pharmacologic Substance Phosphate Buffer Physiological Plasma Plasma Proteins Play Polymerase Chain Reaction Process Property Proteins Protocols documentation RNA vaccine Recovery Recurrence Reproducibility Research Research Personnel Reverse Transcription Role Saline Sampling Silicon Dioxide Students Techniques Technology Temperature Time Uric Acid Virus Work anticancer research aqueous base biomaterial compatibility cancer biomarkers cost digital innovation insight interest microwave electromagnetic radiation parent grant preservation skills solute specific biomarkers undergraduate student user-friendly

项目摘要

Biomarkers play a central role in the detection, diagnosis, management and monitoring of patients with cancer. Factors such as analytical sensitivity and reproducibility are critical to the clinical utility of plasma-based biomarkers in the setting of longitudinal surveillance and early detection of disease and or recurrence. Currently, the state of preservation of biospecimens is accomplished primarily using sub freezer techniques and it is cost prohibitive and certainly not practical in most scenarios. There is clearly a dearth of techniques that can preserve the biospecimens at room temperature for long periods of time. Therefore, there is a critical unmet clinical need for storing a plethora of biospecimens acquired routinely. In the parent grant, we proposed to demonstrate stability and integrity of Pancreatic Cancer Specific biomarkers (CA-A19, miRNA21, and ctDNA) in buffers as well as plasma for extended periods of time at room temperature utilizing a 1-step microwave based sol-gel process. The specific aims were: The specific aims of this research were: (A) Achieve complete integrity and stability of cancer biomarkers in physiologic electrolytes at room temperature. (B) Demonstrate integrity and stability of cancer biomarkers in presence the major plasma proteins (albumin, globulin and fibrinogen). (C) Investigate the integrity of induced cancer biomarkers and native biomarkers present in plasma Solutes such as glucose, uric acid, and other nominal proteins can have an effect on CaRGOS. In the Supplement Grant, Lindsay will develop a reverse engineering approach and will work closely with the PIs and students to directly stabilize biomarkers in plasma. Lindsay started working on this approach and have observed critical challenges including: (i) biocompatibility of the process due to even minimal presence of methanol (ii) possible interference of silica with characterization techniques such as dd PCR. (iii) intensive extraction protocols for extraction of biomarkers from plasma and their compatibility with the silica. These three major hurdles form the basis of the supplement grant, where we will need to advance our fundamental understanding of the nature of interactions of silica precursors with a complex matrix such as plasma. The specific aims of the Supplement grant are: (A) Elimination of methanol from the precursor sol prior to addition of biomarker/plasma. (B) Develop robust protocols to separate silica from the biomarkers at the point of downstream processing During the first year, Lindsay will be an undergraduate researcher and obtain her B.S. in Chemical Engineering. During the second year she will obtain her master’s in engineering enabling her to enrich her skill set in this area. This focused project will allow Lindsay to become an independent and confident researcher in future that aligns with her goals to obtain a MD/PhD.

生物标志物在患者的检测、诊断、管理和监测中发挥着核心作用分析灵敏度和重现性等因素对于临床至关重要。基于血浆的生物标志物在纵向监测和早期检测中的效用目前，生物样本的保存状况已完成。主要使用低温冷冻技术，成本高昂，而且在大多数情况下当然不实用显然缺乏可以在室内保存生物样本的技术。因此，存储一个关键的未满足的临床需求。习惯性获得的大量生物样本。在家长资助中，我们建议证明胰腺癌的稳定性和完整性缓冲液和血浆中的特定生物标志物（CA-A19、miRNA21 和 ctDNA）可用于延长利用基于微波的一步式溶胶-凝胶工艺在室温下放置一段时间。具体目标是：这项研究的具体目标是： (A) 实现癌症的完全完整性和稳定性 (B) 表现出诚信和稳定性主要血浆蛋白（白蛋白、球蛋白和纤维蛋白原）中存在的癌症生物标志物 (C)。研究血浆中存在的诱导癌症生物标志物和天然生物标志物的完整性葡萄糖、尿酸和其他名义蛋白质等溶质会对 CaGOS 产生影响。在补充补助金中，林赛将开发一种逆向工程方法，并将工作 Lindsay 开始与 PI 和学生密切合作，直接稳定血浆中的生物标志物。关于这种方法并观察到了关键挑战，包括：（i）该过程的生物相容性由于即使存在极少量的甲醇 (ii)，二氧化硅也可能干扰表征技术，例如 dd PCR，用于从其中提取生物标志物的强化提取方案。等离子体及其与二氧化硅的相容性这三个主要障碍构成了该技术的基础。补充赠款，我们需要增进对本质的基本理解二氧化硅前体与等离子体等复杂基质的相互作用。补助金为： (A) 在添加生物标记物/血浆之前从前体溶胶中消除甲醇 (B)。开发强大的方案以在下游点将二氧化硅与生物标记物分离加工在第一年，Lindsay 将成为一名本科研究员并获得化学学士学位。在第二年，她将获得工程硕士学位，使她能够这个重点项目将使 Lindsay 成为一名独立的人，丰富她在这一领域的技能。以及对未来充满信心的研究人员，与她获得医学博士/博士学位的目标相一致。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Long-term stabilization of DNA at room temperature using a one-step microwave assisted process.

使用一步微波辅助过程在室温下长期稳定 DNA。

DOI：
发表时间：
2022
期刊：
影响因子：
3.8
作者：
Narvaez Villarrubia, Claudia W;Tumas, Keyla C;Chauhan, Rajat;MacDonald, Thomas;Dattelbaum, Andrew M;Omberg, Kristin;Gupta, Gautam
通讯作者：
Gupta, Gautam

Investigating BioCaRGOS, a Sol-Gel Matrix for the Stability of Heme Proteins under Enzymatic Degradation and Low pH.

研究 BioCaRGOS（一种溶胶-凝胶基质），用于研究酶降解和低 pH 条件下血红素蛋白的稳定性。

DOI：
发表时间：
2023-09-05
期刊：
影响因子：
4.1
作者：
Potnis, Chinmay S;Grapperhaus, Craig A;Gupta, Gautam
通讯作者：
Gupta, Gautam

Long term storage of miRNA at room and elevated temperatures in a silica sol-gel matrix.

在硅溶胶-凝胶基质中在室温和高温下长期储存 miRNA。

DOI：
发表时间：
2021-09-21
期刊：
影响因子：
3.9
作者：
Chauhan, Rajat;Kalbfleisch, Theodore S;Potnis, Chinmay S;Bansal, Meenakshi;Linder, Mark W;Keynton, Robert S;Gupta, Gautam
通讯作者：
Gupta, Gautam

Enhancing the compatibility of BioCaRGOS silica sol-gel technology with ctDNA extraction and droplet digital PCR (ddPCR) analysis.

增强 BioCaRGOS 硅胶溶胶技术与 ctDNA 提取和微滴数字 PCR (ddPCR) 分析的兼容性。

DOI：
发表时间：
2022-10-11
期刊：
影响因子：
3.9
作者：
Potnis, Chinmay S;Chauhan, Rajat;Kalbfleisch, Theodore S;Alexander, Evan;Eichhold, Lindsay;Bansal, Meenakshi;Grapperhaus, Craig A;Keynton, Robert S;Linder, Mark W;Gupta, Gautam
通讯作者：
Gupta, Gautam