Thiroredoxin Targeted Nanoparticles for Cancer Research

用于癌症研究的硫氧还蛋白靶向纳米颗粒

• 批准号: 7845280
• 负责人: STEVEN Sidney SMITH
• 金额: $ 2.06万
• 依托单位: BECKMAN RESEARCH INSTITUTE/CITY OF HOPE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-01 至 2010-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7845280
• 关键词: Aftercare; Antibodies; Back; Binding; Biological Assay; CWR22Rv1; Cells; Chimeric Proteins; Clinical; Color; Custom; DNA; Diagnostic; Diagnostic Procedure; Epithelial Cells; Fluorescein; Fluoresceins; Freezing; Gleason Grade for Prostate Cancer; Glutamate Carboxypeptidase II; Goals; Histology; Home environment; Image; Imagery; Immunohistochemistry; LNCaP; Label; MCF7 cell; Malignant Neoplasms; Malignant neoplasm of prostate; Methyltransferase; Microscope; Nomograms; PC3 cell line; PSA level; Particle Size; Patients; Prostate; Prostatic Neoplasms; Recurrence; Sampling; Staining method; Stains; Stromal Cells; Technology; Testing; Therapeutic; Thioredoxin; Work; abstracting; anticancer research; cancer cell; design; follow-up; improved; nanobiotechnology; nanoparticle; nanoscale; neoplastic cell; research study; scaffold; tool; tumor

DESCRIPTION (provided by applicant): Abstract Currently an important clinical problem in prostate cancer research is predicting the likelihood of recurrence. Our hypothesis is that histology using nanoparticles displaying thioredoxin on tumor sections will improve on the current standard diagnostic methods (e.g. PSA level and Gleason's score) in predicting the likelihood of recurrence. Using this bionanotechnology we have constructed a nanoparticle consisting of a DNA scaffold displaying three copies of the methyltransferase-thioredoxin fusion protein. The DNA scaffold has been modified so as to permit the visualization of the cancer cells using fluorescein. Our preliminary work shows that the nanoparticle displaying thioredoxin binds selectively to LNCaP and MCF-7, but not PC-3, COS-7 and Primary Prostate Epithelial Cells, suggesting that the nanoparticle is selectively targeting certain types of cancer cells. We propose to follow up on these preliminary findings with the following Specific Aims: Specific Aim 1: (i) Generate nanoparticles comprising a Y-junction DNA scaffold displaying three identical methyltransferase thioredoxin fusion proteins (YRII-Trx-F) in quantities necessary for the experiments described in Specific Aims 2 and 3. (ii) Generate and purify thioredoxin labeled directly with fluorescein (Trx-F) as a control. Specific Aim 2: Test the nanoparticle displaying the thioredoxin (YRII-Trx-F) and thioredoxin labeled directly with fluorescein (Trx-F) for their ability to discriminate between various available prostate cancer cell lines: (e.g. LNCaP, PC-3, DU-145, MDA PCa 2b and 22Rv1) while not detecting primary prostate stromal cells, and primary prostate epithelial cells. Specific Aim 3: Test the nanoparticle displaying the thioredoxin (YRII-Trx-F) and thioredoxin labeled directly with fluorescein (Trx-F) determine whether or not the binding to frozen prostate tumor sections can be correlated with Gleason's score and/or recurrence. For comparison, immunohistochemistry will be performed on sections of each tumor sample to evaluate the level of expression of PSMA and AMACR using anti-PSMA antibodies and and-AMACR antibodies. Each diagnostic assay will be analyzed to see if there is a correlation between the Gleason's Sum, antibody binding intensity and fluorescent intensity of the nanoparticles. Our long-term goals are two-fold: First, we hope that our findings will provide a new parameter that can contribute to a nomogram for the prediction of recurrence. Second, the modular design of the nanoparticle allows us to custom design it to target almost any cancer for potential delivery into cells. If the results of this study show that this nanoparticle can specifically target aggressive prostate tumor cells, we could then modify the design for potential delivery of a therapeutic into the cells. Both goals are beyond the scope of the present proposal. Project Narrative Currently our tools for determining who is in danger of having prostate cancer return are not as helpful as they should be, so we cannot follow up with additional treatment until the cancer reappears. We have developed a technology for creating nanoscale (molecule size) particles that can home in on the cancer cells and make them glow. In this proposal we will attempt to determine whether or not the color we see under the microscope will permit us to pick out those cancers that are likely to come back after treatment and help the patients who are likely to come down with the cancer again to take additional action.

描述（由申请人提供）：摘要目前，前列腺癌研究中一个重要的临床问题预测了复发的可能性。我们的假设是，在肿瘤切片上显示硫氧还蛋白的纳米颗粒的组织学将改善当前标准诊断方法（例如，PSA水平和格里森的得分），以预测复发的可能性。使用这种Bionanotechnology，我们构建了由DNA支架组成的纳米颗粒，该纳米颗粒显示了三个副本的甲基转移酶 - 硫代蛋白融合蛋白。 DNA支架已被修饰，以便使用荧光素对癌细胞的可视化。我们的初步工作表明，表现出硫氧还蛋白的纳米颗粒与LNCAP和MCF-7有选择性结合，但不选择PC-3，COS-7和原代前列腺上皮细胞，这表明纳米颗粒选择性地靶向某些类型的癌细胞。我们建议对这些初步发现进行以下特定目的进行跟进：（i）生成纳米颗粒，其中包含Y结DNA支架，显示三个相同的甲基转移酶硫代蛋白融合蛋白融合蛋白（YRII-I-TRX-F）在特定目标中所描述的实验所需的数量所需的数量所必需的三个相同的甲基转移蛋白支架。荧光素（TRX-F）作为对照。 Specific Aim 2: Test the nanoparticle displaying the thioredoxin (YRII-Trx-F) and thioredoxin labeled directly with fluorescein (Trx-F) for their ability to discriminate between various available prostate cancer cell lines: (e.g. LNCaP, PC-3, DU-145, MDA PCa 2b and 22Rv1) while not detecting primary prostate stromal cells, and primary prostate上皮细胞。具体目标3：测试显示硫氧还蛋白（YRII-TRX-F）和硫氧还蛋白直接标记的硫蛋白（TRX-F）标记的纳米颗粒确定与GLEASON的分数和/或复发的结合是否可以与冻结前列腺肿瘤切片的结合相关。为了进行比较，将在每个肿瘤样品的切片上进行免疫组织化学，以使用抗PSMA抗体和AMACR抗体评估PSMA和AMACR的表达水平。将分析每个诊断测定法，以查看格里森总和，抗体结合强度和纳米颗粒的荧光强度之间是否存在相关性。我们的长期目标是两个方面：首先，我们希望我们的发现将提供一个新的参数，可以为复发预测的诺夫图做出贡献。其次，纳米颗粒的模块化设计使我们能够将其自定义，以将其靶向几乎所有癌症，以供潜在的细胞递送。如果这项研究的结果表明该纳米颗粒可以特异性地靶向侵袭性的前列腺肿瘤细胞，则可以修改设计的设计，以便将治疗性递送到细胞中。这两个目标都超出了本提议的范围。目前，我们确定谁有前列腺癌返回的危险的工具叙述不像应有的帮助，因此直到癌症再次出现，我们才能进行其他治疗。我们已经开发了一种创建可以在癌细胞中回家并使其发光的纳米级（分子大小）颗粒的技术。在此提案中，我们将尝试确定我们在显微镜下看到的颜色是否允许我们挑选出可能在治疗后可能会回来的那些癌症，并帮助可能再次患上癌症的患者采取其他行动。

项目成果

The influence of PSA-RNA yield on the analysis of expressed prostatic secretions (EPS) for prostate cancer diagnosis.

• 发表时间: 2013-02
• 期刊: The Canadian journal of urology
• 作者: C. Whelan;L. Crocitto;M. Kawachi;Kevin G. Chan;David D. Smith;T. Wilson;Steven M. Smith

2'-Deoxyriboguanylurea, the primary breakdown product of 5-aza-2'-deoxyribocytidine, is a mutagen, an epimutagen, an inhibitor of DNA methyltransferases and an inducer of 5-azacytidine-type fragile sites.

2-Deoxyriboguanylurea 是 5-aza-2-deoxyribocytidine 的主要分解产物，是一种诱变剂、表观诱变剂、DNA 甲基转移酶抑制剂和 5-azacytidine 型脆弱位点诱导剂。

• DOI: 10.1093/nar/gks706
• 发表时间: 2012
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Lamparska,Katarzyna;Clark,Jarrod;Babilonia,Gail;Bedell,Victoria;Yip,Wesley;Smith,StevenS
• 通讯作者: Smith,StevenS

Expressed prostatic secretion biomarkers improve stratification of NCCN active surveillance candidates: performance of secretion capacity and TMPRSS2:ERG models.

表达的前列腺分泌生物标志物改善了 NCCN 主动监测候选者的分层：分泌能力和 TMPRSS2:ERG 模型的表现。

• DOI: 10.1016/j.juro.2013.05.019
• 发表时间: 2014
• 期刊: The Journal of urology
• 作者: Whelan,Christopher;Kawachi,Mark;Smith,DavidD;Linehan,Jennifer;Babilonia,Gail;Mejia,Rosa;Wilson,Timothy;Smith,StevenS
• 通讯作者: Smith,StevenS

Presurgical Biomarker Performance in the Detection of Gleason Upgrading in Prostate Cancer.

• DOI: 10.1158/1055-9965.epi-16-0488
• 发表时间: 2016-12
• 期刊: Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology
• 作者: Wittig K;Yamzon JL;Smith DD;Jeske DR;Smith SS
• 通讯作者: Smith SS

Human non-CG methylation: are human stem cells plant-like?

• DOI: 10.4161/epi.5.7.12702
• 发表时间: 2010-10-01
• 期刊: Epigenetics
• 影响因子: 3.7
• 作者: Dyachenko OV;Schevchuk TV;Kretzner L;Buryanov YI;Smith SS
• 通讯作者: Smith SS