前列腺癌雄激素受体功能的预后和治疗分层评价
Although most prostate cancers are initially responsive to androgen ablation therapy, they become treatment resistant as tumor cells develop mechanisms to evade the treatment. Early knowledge of the androgen receptor dysfunctions will help in patient stratification for emerging therapeutic strategies. We proposed an approach for monitoring potential dysfunctions of the androgen receptor by measuring expression of a panel of genes directly regulated by androgen receptor. We examined human prostate cancer tissues (surgery or diagnostic biopsy specimens) at early stages of the disease and matched with longitudinal follow up data. Within the first reporting period we have completed the quality control of detecting PSA/KLK3, PMEPA1, NKX3.1, ODC1, AMD1 and TMPRSS2-ERG genes in VCap cell culture model monitoring kinetic and dose response to androgen. In the second reporting period we have completed the qRT-PCR evaluation of in 77 patients by monitoring ERG, PSA, PMEPA1 and GAPDH levels. Also, we have completed the evaluation of 40 whole mounted sections of RP specimens by immunohistochemistry assessing AR, ERG, NKX3.1 and PSA proteins and compared the results to corresponding GeneCHip mRNA expression levels from the same tumor foci. The result indicated remarkable accuracy of the androgen regulated gene expression at mRNA levels performed better in prediction favorable outcomes in tumors with well differentiated morphology. By the completion of the proposed research we will provide a quantitative index of AR dysfunction for enhancing prognostic accuracy and to stratify patients for specific therapeutic approaches at early stages of prostate cancer treatment.
蛋白质组装的魔角旋转NMR研究:方法及应用的最新进展
In living organisms, many processes are executed through tightly regulated networks of multi-component macromolecular complexes. Knowledge of structure and dynamics of such assemblies is essential to understanding their biological mechanisms. Solid-state NMR spectroscopy is a prime method for studying macromolecular assemblies, at atomic-level resolution, particularly in systems that are insoluble and lack long-range order. In this report, we review the contemporary magic-angle-spinning (MAS) NMR methods for investigation of proteins assemblies. We describe the progress in multidimensional correlation spectroscopy, recoupling, and decoupling techniques. We discuss emerging methods for recording spectra at MAS frequencies of 40 kHz and faster. We review techniques for sensitivity enhancement and fast data collection. We address isotopic labelling approaches to alleviate spectral congestion, facilitate assignments, and highlight specific intra- and intermolecular interactions. Finally, we present recent case studies on exciting and challenging systems, including assemblies of bacterio-phages and HIV-1 proteins, cytoskeleton-associated proteins, gas vesicles, type Ⅲ secretion system, and protein-RNA complexes.