20 research outputs found
An informatics model for tissue banks – Lessons learned from the Cooperative Prostate Cancer Tissue Resource
BACKGROUND: Advances in molecular biology and growing requirements from biomarker validation studies have generated a need for tissue banks to provide quality-controlled tissue samples with standardized clinical annotation. The NCI Cooperative Prostate Cancer Tissue Resource (CPCTR) is a distributed tissue bank that comprises four academic centers and provides thousands of clinically annotated prostate cancer specimens to researchers. Here we describe the CPCTR information management system architecture, common data element (CDE) development, query interfaces, data curation, and quality control. METHODS: Data managers review the medical records to collect and continuously update information for the 145 clinical, pathological and inventorial CDEs that the Resource maintains for each case. An Access-based data entry tool provides de-identification and a standard communication mechanism between each group and a central CPCTR database. Standardized automated quality control audits have been implemented. Centrally, an Oracle database has web interfaces allowing multiple user-types, including the general public, to mine de-identified information from all of the sites with three levels of specificity and granularity as well as to request tissues through a formal letter of intent. RESULTS: Since July 2003, CPCTR has offered over 6,000 cases (38,000 blocks) of highly characterized prostate cancer biospecimens, including several tissue microarrays (TMA). The Resource developed a website with interfaces for the general public as well as researchers and internal members. These user groups have utilized the web-tools for public query of summary data on the cases that were available, to prepare requests, and to receive tissues. As of December 2005, the Resource received over 130 tissue requests, of which 45 have been reviewed, approved and filled. Additionally, the Resource implemented the TMA Data Exchange Specification in its TMA program and created a computer program for calculating PSA recurrence. CONCLUSION: Building a biorepository infrastructure that meets today's research needs involves time and input of many individuals from diverse disciplines. The CPCTR can provide large volumes of carefully annotated prostate tissue for research initiatives such as Specialized Programs of Research Excellence (SPOREs) and for biomarker validation studies and its experience can help development of collaborative, large scale, virtual tissue banks in other organ systems
Examination of the Fractalkine and Fractalkine Receptor Expression in Fallopian Adenocarcinoma Reveals Differences When Compared to Ovarian Carcinoma
Fallopian adenocarcinoma is a rare malignancy arising in the epithelium of the fallopian tube. Fallopian tube epithelium has been proposed as a tissue origin for high-grade serous ovarian carcinoma, the deadliest gynecologic malignancy. Given the commonalities in dissemination and treatment of these malignancies, we contemplated the possibility of similar patterns of gene expression underlying their progression. To reveal potential similarities or differences in the gene expression of fallopian adenocarcinoma and high-grade serous ovarian carcinoma, we tested expression of the fractalkine receptor (CX3CR1) and its ligand, fractalkine (CX3CL1), in the specimens of normal and pathologic fallopian tube using immunohistochemistry. Our data show that CX3CR1 is expressed in the normal, cancer adjacent normal, inflammatory, and malignant fallopian epithelium. CX3CL1 was expressed only by the normal and cancer adjacent normal fallopian tube epithelium; its expression was largely lost in the inflammatory and malignant fallopian epithelium. In opposite, both CX3CR1 and CX3CL1 are expressed in high-grade serous ovarian carcinoma. These findings are consistent with an idea that fallopian adenocarcinoma and high-grade serous ovarian carcinoma, although currently thought to arise from the same organ, may not share similar molecular characteristics
Three-Dimensional Collagen Type I Matrix Up-Regulates Nuclear Isoforms of the Microtubule Associated Protein Tau Implicated in Resistance to Paclitaxel Therapy in Ovarian Carcinoma
Epithelial ovarian carcinoma is the deadliest gynecologic malignancy. One reason underlying treatment failure is resistance to paclitaxel. Expression of the microtubule associated protein tau has recently been proposed as a predictor of response to paclitaxel in ovarian carcinoma patients. Expression of tau was probed using immunohistochemistry in 312 specimens of primary, and 40 specimens of metastatic, ovarian carcinoma. Serous epithelial ovarian carcinoma cell line models were used to determine the expression of tau by Western blot and immunofluorescence staining. Subcellular fractionation and Western blot were employed to examine nuclear and cytoplasmic localization of tau. Gene silencing and clonogenic assays were used to evaluate paclitaxel response. Tau was expressed in 44% of all tested cases. Among the primary serous epithelial ovarian carcinoma cases, 46% were tau-positive. Among the metastatic serous epithelial ovarian carcinomas, 63% were tau-positive. Cell culture experiments demonstrated that tau was expressed in multiple isoforms. Three-dimensional collagen I matrix culture conditions resulted in up-regulation of tau protein. Silencing of tau with specific siRNAs in a combination with three-dimensional culture conditions led to a significant decrease of the clonogenic ability of cells treated with paclitaxel. The data suggest that reduction of tau expression may sensitize ovarian carcinoma to the paclitaxel treatment
Analysis of iron, zinc, selenium and cadmium in paraffin-embedded prostate tissue specimens using inductively coupled plasma mass-spectrometry
Formalin-fixed paraffin-embedded (FFPE) tissue specimens represent a valuable and abundant resource of pathologic material for various biomedical studies. In the present study, we report the application of high-resolution inductively coupled mass-spectrometry (ICP-MS) for quantification of Fe, Zn, Se and Cd in FFPE prostate tissue. These elements have a possible role in the development of prostate diseases: while Zn and Se are needed for a healthy prostate, Cd shows multiple toxic and carcinogenic effects. Excessive accumulation of Fe induces the production of highly reactive hydroxyl radical species, which may play a role in cancer etiopathogenesis. To assess whether the levels of these metals in the FFPE prostate tissue represent their original content, we compared their levels with those in the fresh tissue (on dry weight basis) in samples obtained from 15 patients. We found that in FFPE tissue, the recoveries of Se, Fe, Cd and Zn were progressively decreased, 97±11% (r=0.88), 82±22% (r=0.86), 59±23% (r=0.69) and 24±11% (r=0.38), respectively. Thus, the use of correction factors, determined as k=0.16 for Se, k=0.20 for Fe, k=0.27 for Cd and k=0.67 for Zn, is required to estimate the retrospective levels of these elements in the parental non-processed fresh (wet) prostate tissue. The technique used in this study enables the analysis of archival FFPE prostate tissue for the concentrations of Fe, Zn, Se and Cd to study association between the levels of these metals and prostate disease
Fractalkine receptor is expressed in mature ovarian teratomas and required for epidermal lineage differentiation
BACKGROUND:
The goal of this study was to determine a predominant cell type expressing fractalkine receptor (CX3CR1) in mature ovarian teratomas and to establish functional significance of its expression in cell differentiation.
METHODS:
Specimens of ovarian teratoma and human fetal tissues were analyzed by immunohistochemistry for CX3CR1expression. Ovarian teratocarcinoma cell line PA-1 was used as a model for cell differentiation.
RESULTS:
We found that the majority of the specimens contained CX3CR1-positive cells of epidermal lineage. Skin keratinocytes in fetal tissues were also CX3CR1- positive. PA-1 cells with downregulated CX3CR1 failed to express a skin keratinocyte marker cytokeratin 14 when cultured on Matrigel in the presence of a morphogen, bone morphogenic protein 4 (BMP-4), as compared to those expressing scrambled shRNA.
CONCLUSIONS:
Here we demonstrate that CX3CR1 is expressed in both normally (fetal skin) and abnormally (ovarian teratoma) differentiated keratinocytes and is required for cell differentiation into epidermal lineage
Prostate cancer outcome and tissue levels of metal ions
Background: There are many studies on prostate cancer incidence and exposure to heavy metals such as cadmium, iron, selenium and zinc. Much less data is available on the possible influence of heavy metals on prostate cancer outcome. We and others have shown that cadmium, iron and zinc exposure in vitro can cause changes in cell invasion. Recently we have published an inductively coupled plasma-mass spectrometry (ICP-MS) method to reliably measure concentration of these metals in paraffin embedded archival tissues. Here we study the level of these ions in the prostate of patients with known outcome in order to address metals and disease progression.
Methods: The source of tissues Cooperative Prostate Cancer Tissue Resource (CPCTR). We obtained formalin fixed paraffin embedded tissue blocks of prostatectomy samples of 40 patients with PSA recurrence, matched 1:1 (for year of surgery, race, age, Gleason grading score and stage) with tissue blocks from 40 patients without recurrence (n=80). All non-recurrence patients had more than 5-years post-prostatectomy follow-up. Case-control pairs were compared for the levels of heavy metals in areas adjacent to tumors. After deparaffinization, each tissue sample was oven-dried and divided in 5-10 µg triplicates. Internal standards were added (Y and 74Se), digested overnight in nitric acid. ICP-MS was performed by comparing samples with standard solutions of each analyte (Cd, Fe, Zn, and Se). The association of each individual metal with prostate cancer recurrence was evaluated by Wilcoxon signed rank paired test comparing matched recurrence cases and non-recurrence control cases.
Results: Patients with biochemical (PSA) recurrence of disease had 14% lower iron [95 vs. 112 parts per million (ppm); p = 0.04] and 9% lower zinc (28 vs. 35 ppm; p = 0.04) concentrations in the normal-appearing tissue immediately adjacent to prostate cancer areas. Differences in cadmium [488 vs. 439 parts per billion (ppb); 10% higher) and selenium (1676 vs. 1576 ppb; 6% higher] levels were not statistically significant in recurrence cases when compared to non-recurrences (p=0.40 and 0.21 respectively).
Discussion: To the best of our knowledge this is the first study to show an association between metal tissue levels and outcomes in prostate cancer. Lower zinc in cases of worse outcome suggests this may be an area for further investigation in recurrence chemoprevention. Lower iron may be the result of iron sequestration in the reticuloendothelial system caused by aggressive cancer, rather than a factor important for prostate cancer progression, a hypothesis that needs further investigation. The lack of association of recurrence with either cadmium or selenium was an unexpected finding
Prostate cancer outcome and tissue levels of metal ions
Background: There are many studies on prostate cancer incidence and exposure to heavy metals such as cadmium, iron, selenium and zinc. Much less data is available on the possible influence of heavy metals on prostate cancer outcome. We and others have shown that cadmium, iron and zinc exposure in vitro can cause changes in cell invasion. Recently we have published an inductively coupled plasma-mass spectrometry (ICP-MS) method to reliably measure concentration of these metals in paraffin embedded archival tissues. Here we study the level of these ions in the prostate of patients with known outcome in order to address metals and disease progression. \ud
\ud
Methods: The source of tissues Cooperative Prostate Cancer Tissue Resource (CPCTR). We obtained formalin fixed paraffin embedded tissue blocks of prostatectomy samples of 40 patients with PSA recurrence, matched 1:1 (for year of surgery, race, age, Gleason grading score and stage) with tissue blocks from 40 patients without recurrence (n=80). All non-recurrence patients had more than 5-years post-prostatectomy follow-up. Case-control pairs were compared for the levels of heavy metals in areas adjacent to tumors. After deparaffinization, each tissue sample was oven-dried and divided in 5-10 µg triplicates. Internal standards were added (Y and 74Se), digested overnight in nitric acid. ICP-MS was performed by comparing samples with standard solutions of each analyte (Cd, Fe, Zn, and Se). The association of each individual metal with prostate cancer recurrence was evaluated by Wilcoxon signed rank paired test comparing matched recurrence cases and non-recurrence control cases. \ud
\ud
Results: Patients with biochemical (PSA) recurrence of disease had 14% lower iron [95 vs. 112 parts per million (ppm); p = 0.04] and 9% lower zinc (28 vs. 35 ppm; p = 0.04) concentrations in the normal-appearing tissue immediately adjacent to prostate cancer areas. Differences in cadmium [488 vs. 439 parts per billion (ppb); 10% higher) and selenium (1676 vs. 1576 ppb; 6% higher] levels were not statistically significant in recurrence cases when compared to non-recurrences (p=0.40 and 0.21 respectively). \ud
\ud
Discussion: To the best of our knowledge this is the first study to show an association between metal tissue levels and outcomes in prostate cancer. Lower zinc in cases of worse outcome suggests this may be an area for further investigation in recurrence chemoprevention. Lower iron may be the result of iron sequestration in the reticuloendothelial system caused by aggressive cancer, rather than a factor important for prostate cancer progression, a hypothesis that needs further investigation. The lack of association of recurrence with either cadmium or selenium was an unexpected finding