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Hans Kreipe

Publications and source records attributed to Hans Kreipe.

69 records · Page 4Linked to original sources
PubMed2003

Rapid and large-scale transition of new tumor biomarkers to clinical biopsy material by innovative tissue microarray systems.

A rapidly increasing number of studies are being published providing significant results by the use of tissue microarrays (TMAs), ie, paraffin blocks composed of multiple specimens. The numerous advantages of this technology are obvious and have thus stimulated many constructors to evolve and improve different technical approaches. With TMAs, multiple specimens can be simultaneously investigated with different in situ techniques under identical laboratory conditions, resulting in a dramatic time and cost reduction compared with conventional pathologic studies. Furthermore, this technology is less exhausting for the finite original donor material, allowing for a significantly increased number of assays per each case. Against the background of decoding the human genome and the widespread application of high-density cDNA microarrays, the next challenge will be to apply the genome discoveries to the clinical setting. For pathologists, TMAs represent an ideal adjunct and can be very useful for the rapid and high-throughput discovery and validation of countless candidate biomarkers, assessing their prognostic and predictive value and identifying further therapy targets. This article provides a review of different TMA technologies and addresses the technical aspects of their construction and their validity in different applications through data from the literature along with the authors' own experiences.

Biomarkers, Tumor↗
PubMed2003

Evolution of myelofibrosis in chronic idiopathic myelofibrosis as evidenced in sequential bone marrow biopsy specimens.

Although the new World Health Organization (WHO) classification acknowledges "prefibrotic" phases, progression of myelofibrosis in chronic idiopathic myelofibrosis (cIMF) is controversial because there are only a few studies about sequential biopsy specimens, and they yield conflicting results. The conflicting results might be due to a mixture of different degrees of myelofibrosis and therapy regimens within the respective groups studied. To prove this hypothesis, we studied sequential bone marrow biopsy specimens from patients with cIMF and compared 3 groups with different degrees of myelofibrosis at initial diagnosis with a group of patients with primarily unfibrosed disease who met the WHO criteria for prefibrotic cIMF. Patients receiving chemotherapy were considered separately from patients without treatment. Our results favor a steady progression of myelofibrosis unrelated to therapy modalities, whereas confusing literature data can be explained: fibrosis may remain static or lessen, especially in more advanced stages of cIMF.

Antineoplastic Agents↗
PubMed2002

Promoter hypermethylation of the death-associated protein kinase gene in breast cancer is associated with the invasive lobular subtype.

Expression of death-associated protein (DAP) kinase, a proapoptotic serine/threonine protein kinase, is frequently lost in human tumors. In a study of 134 primary breast cancer specimens hypermethylation of the DAP kinase gene was found in 13% of cases. A highly significant difference (P < 0.001) of DAP kinase inactivation was observed between invasive lobular cancer (n = 19) and invasive ductal cancer (n = 85; 53% versus 9%, respectively). Hypermethylation correlated with loss of RNA expression, estrogen receptor positivity (P < 0.01), and the absence of p53 overexpression (P < 0.01). In contrast to invasive lobular cancer, the in situ-growing precursor lesion lacked epigenetic modification of the DAP kinase promotor by aberrant methylation indicating a potential role in tumor progression. Unlike the DAP kinase gene, hypermethylation of the cyclin D2 and RASSF1A genes did not correlate with a particular histological subtype or to invasiveness [corrected]. We conclude that different histological subtypes of breast cancer may not only differ concerning specific chromosomal abnormalities and DNA mutations but also with regard to epigenetic inactivation patterns.

Apoptosis Regulatory Proteins↗
PubMed2002

Inter-laboratory and inter-observer reproducibility of immunohistochemical assessment of the Ki-67 labelling index in a large multi-centre trial.

Proliferative activity of tumour cells, as assessed by the Ki-67 labelling index, has been suggested as a potential prognostic indicator in many neoplastic diseases. Meaningful application of the immunohistochemically determined tumour cell growth fraction in clinical decision-making requires information about its inter-laboratory reproducibility. To assess the reproducibility of Ki-67 determined growth fraction, a multi-centre immunohistochemical trial was performed with 172 participating laboratories, each testing 30 different tissue samples. Evaluating 5160 Ki-67 labelling indices with a newly developed tissue microarray, good inter-observer reproducibility but high inter-laboratory variability was found. Reassessment of all stainings revealed considerable inter-laboratory differences in the intensity and frequency of labelled nuclei, suggesting that antigen retrieval or staining techniques are predominantly responsible for the inter-laboratory variability found in this trial. Consequently, cut-off levels for Ki-67, suggested to distinguish prognostic subgroups in tumours, appear to have limited reproducibility in a multi-centre approach. It is concluded that there is a need to standardize the immunohistochemical determination of the Ki-67 labelling index when it is used as a prognostic indicator in surgical pathology.

Biomarkers, Tumor↗
PubMed2002

Comparative genomic hybridization (CGH) and fluorescence in situ hybridization (FISH) in the diagnosis of hepatocellular carcinoma.

BACKGROUND: The histomorphological diagnosis of well differentiated hepatocellular carcinoma (HCC) may be a challenging task, because regenerative nodules and adenomas share similar microscopic features. Moreover, the differentiation of intrahepatic metastatic spread from multicentric growth in multinodular HCC is frequently not possible by histological criteria alone. Whether molecular cytogenetic analysis can contribute to the differential diagnosis of HCC was recently investigated by our group. METHODS: We applied comparative genomic hybridization (CGH) and fluorescence in situ hybridization (FISH) to 12 cases of hepatocellular adenoma (HCA) and 28 cases of well differentiated HCC. RESULTS: CGH revealed aberrations in 2/6 HCAs, affecting 7p in 1 sample each and 17q and 20 in the second case. In 4/4 well differentiated HCCs, aberrations occurred, with a mean of eight aberrations per case, focused on 1q, 4q, 8p, 8q, 16p, and 17p. In all HCC samples, at least two of these sites were affected. In 2 multinodular HCCs, six and four different tumor nodes, respectively, were analyzed. In the first case, aberration patterns of chromosomes 1, 4, 5, 9, 13, and 17 were almost identical in all six nodes, indicating metastatic spread. In the second case, three of the nodes showed very similar patterns of chromosome aberrations, including those of chromosomes 1, 4, 5, 7, 8, 10, 12, 14, 17, and 18. Aberrations of chromosomes 4, 5, 8, 10, 12, and 15 were shown in the fourth node, findings conclusive for both the metastatic spread and the multicentric origin of the HCC. Based on the CGH results, five probes, for chromosomes 1, 6, 7, 8, and X, were selected for FISH. Using this panel, we found no aberrations in 14 HCAs. By contrast, 13/14 HCCs demonstrated aberrations for two to five chromosomes in the FISH analysis. CONCLUSIONS: We conclude that CGH and FISH are helpful diagnostic tools for the histopathological differential diagnosis of HCC.

Adenoma, Liver Cell↗
PubMed2002

Quantitative assessment of promoter hypermethylation during breast cancer development.

The aberrant methylation of cytosine residues in the promoter region of growth regulatory genes is now widely recognized as an additional mechanism for gene inactivation in cancer cells. In this study we analyzed the methylation status of four growth regulatory genes (p16, RASSF1A, cyclinD2, 14-3-3zeta) during breast cancer progression. For this purpose invasive and noninvasive tumor cell populations as well as hyperplastic cell proliferations were isolated from a series of archival breast tissue specimens (n = 57) using laser-assisted microdissection. A new real-time polymerase chain reaction-based assay was used for the sensitive and quantitative determination of the cell-specific methylation status. We found that aberrant promoter methylation was already prevalent in pure intraductal carcinoma with different frequencies and different methylation levels for the four genes analyzed. For RASSF1A and 14-3-3zeta promoter methylation was also demonstrated in epithelial hyperplasia and intraductal papillomas. By contrast, aberrant methylation of cyclinD2 and p16 was restricted to cancerous epithelium. Increased methylation of the cyclinD2 gene was significantly associated with a higher van Nuys grade. Furthermore, when intraductal and invasive tumor cells were compared, significant quantitative changes in the methylation level were detected primarily within the cyclinD2 gene. These results demonstrate that promoter methylation is an early and frequent event in breast cancer development, but displays great quantitative and gene-specific differences, and changes in a gene-specific manner during tumor progression.

14-3-3 Proteins↗
PubMed2002

Detection of chromosomal aberrations in well-differentiated hepatocellular carcinoma by bright-field in situ hybridization.

Differentiation between well-differentiated hepatocellular carcinoma (HCC) and nonmalignant lesions with increased cellular proliferation may be difficult in needle biopsies. Based on recurrent chromosome aberrations known for HCC, we developed a nonfluorescent in situ hybridization technique that allows combination with morphological analysis in bright-field microscopy. Fourteen biopsies of HCC and 31 samples of regenerative nodules (n = 10), chronic hepatitis (n = 10), fibrosis or cirrhosis of unknown origin (n = 5), focal nodular hyperplasia (n = 2), primary biliary cirrhosis (n = 2), steatosis (n = 1), and adenomatous hyperplasia (n = 1) were analyzed with probes specific for the centromeric regions of chromosomes 1, 6, 7, and 8. After microwave pretreatment and in situ hybridization, signals were detected using a tyramine-based system and AEC as substrate. Evaluation of signals was done by conventional bright-field microscopy. Using this approach, aberrant counts were seen for at least one chromosome in 12/14 cases of HCC. In contrast, none of the nonmalignant lesions revealed aberrant counts for any of the chromosomes analyzed. In conclusion, this new combination of in situ hybridization and tyramine amplification allows fast and reliable evaluation of chromosome aberrations in a histomorphological context similar to paraffin immunohistochemistry. Registration of imbalances contributes to a reliable differentiation between malignant and nonmalignant lesions of the liver.

Adult↗
PubMed2002

Reliability of lymphoma classification in bone marrow trephines.

The aim of this study was to test and establish the accuracy and reliability of lymphoma classification in bone marrow trephines according to the new World Health Organization (WHO) classification by considering predominantly the morphology and immunophenotype. Therefore, we retrospectively compared lymphoma diagnoses, rendered exclusively on bone marrow trephines without knowledge of lymph node diagnosis in 124 patients, with the results of the reference centres that had reviewed lymph node (n = 90) or extranodal biopsies (n = 34). The overall concordance rate was higher than 85% and 91%, respectively, when patients with discordant malignancy grades were excluded. The concordance rate for low-grade B-cell lymphomas was 93% and for high-grade B-cell lymphomas 84%. The main reasons for discordant diagnoses were divergent immunophenotypes among low-grade B-cell lymphomas (6 out of 81, i.e. 7.4%) and discrepant malignancy grades within high-grade B-cell lymphomas (6 out of 31, i.e. 19.4%). No relationship between discordant diagnoses and chemotherapy given during the course of the disease with the site of biopsy (i.e. lymph nodes, extranodal sites) was noted. We conclude from our results that bone marrow trephines are a reliable tool, not only for establishing bone marrow infiltration, but also for the subtyping of lymphomas.

Biopsy↗
PubMed2002

Prognostic significance of calcium-binding protein S100A4 in colorectal cancer.

BACKGROUND & AIMS: Prognostication in colon cancer almost exclusively still rests on the tumor stage. Furthermore, tumor-derived markers to improve discrimination of low- and high-risk subtypes generally are not in use. S100A4 has been reported to be associated with invasion and metastasis; however, no data are available on its prognostic value in colorectal carcinoma. Therefore, we investigated the prognostic significance of immunohistochemical S100A4 expression in colorectal carcinoma compared with clinicopathologic parameters and expression of cell-cycle markers p16, p21, p27, p53, Ki-67, and RB. METHODS: Archival tissue from 709 patients with colorectal cancer were retrieved, applied in tissue array technology, and investigated immunohistochemically. Univariate and multivariate survival analyses were carried out on all investigated parameters. RESULTS: Sixteen percent of cases showed high; 31%, low; and 53%, no S100A4 expression. In Kaplan-Meier analysis, S100A4 positively stained cases showed a significantly decreased survival time compared with negatively stained cases (P < 0.0001). In multivariate regression analysis, S100A4 expression emerged as a highly significant independent parameter (P < 0.001) with the highest relative-risk factor among other covariates. Nodal status (pN) lost its prognostic value if S100A4 was added to the model. High S100A4 expression was associated with tumor stage pT3/4, secondary metastasis, women, p16, and RB expression. CONCLUSIONS: S100A4 expression represents a highly significant prognostic marker in colorectal carcinoma, which is able to identify a subset of patients at high risk. In this respect, it is superior to established prognostic markers such as nodal status, pT stage, and p53 expression.

Colorectal Neoplasms↗
PubMed2002

High frequency of epithelial chimerism in liver transplants demonstrated by microdissection and STR-analysis.

It has recently been shown that epithelial cells derived from stem cells originating outside the liver are integrated into liver allografts. Whether epithelial intragraft chimerism protects transplants from rejection or chronic transplant dysfunction, and whether it interferes with recurrence of primary liver disease, is not known. Twenty-seven sequential biopsies derived from 9 liver-transplant recipients were studied for chimerism of hepatocytes and cholangiocytes. The target cells were isolated by laser microdissection after cytokeratin immunolabeling and genotyped using DNA analysis of a highly polymorphic short tandem repeat. Irrespective of whether early (up to 4 weeks) or late (more than 12 months) posttransplantation biopsies were studied, cholangiocyte chimerism was almost constantly found in 91% of the samples. No significant differences occurred between samples derived from patients with chronic organ dysfunction (n = 3), recurrent hepatitis (n = 3), or mild, unspecific changes (n = 3). By contrast, hepatocyte chimerism tended to occur later (55% vs. 22%) and appeared to be associated with recurrent hepatitis (67% vs. 27%). In this respect, chronic organ dysfunction did not differ from mild, unspecific changes. While cholangiocyte chimerism represents a constant and early phenomenon in liver transplantations, an enhanced chimeric integration of recipient-derived hepatocytes can be observed in recurrent hepatitis, supporting the concept of an increased recruitment of extrahepatic progenitor cells to the liver in chronic hepatitis.

Bile Ducts↗
PubMed2002

Marked intratumoral heterogeneity of c-myc and cyclinD1 but not of c-erbB2 amplification in breast cancer.

Intratumoral heterogeneity mirrors subclonal diversity and might affect treatment response. To investigate molecular heterogeneity of primary breast cancer specimens, we determined the amplification status of growth regulatory genes (c-erbB2, topoisomerase IIalpha, c-myc, and cyclinD1) in macroscopically and microscopically separate areas of individual tumors (n = 21). Using laser-assisted microdissection and quantitative PCR, we found marked intratumoral heterogeneity with different patterns for each gene. Molecular heterogeneity in amplification pattern could be demonstrated between both macroscopically (0.5 to several centimeters) and microscopically (10 to several hundred micrometers) distant tumor areas. C-erbB2 amplification proved to be the most stable amplification in individual tumors, with heterogeneity occurring in only 36% of amplified cases. By contrast, amplification of c-myc and cyclinD1 revealed varying patterns in the vast majority of amplified cases (100% and 83%). The constancy of c-erbB2 amplification underlines its presumed importance in breast cancer biology. We conclude that the molecular heterogeneity of breast cancer as evidenced in this study requires thorough and representative sampling of different tumor areas when the biologic significance of somatic mutations is considered. Patterns of heterogeneity can be used to trace the clonal evolution within different compartments of an individual tumor.

Breast Neoplasms↗
PubMed2002

Tissue array technology for testing interlaboratory and interobserver reproducibility of immunohistochemical estrogen receptor analysis in a large multicenter trial.

Semiquantitative immunohistochemical assessment of estrogen receptor (ER) is used to predict the likelihood of response to antiestrogen therapy in breast carcinoma. If semiquantitative immunohistochemical analysis leads to therapeutic decisions, the importance of standardization and quality control increases. ER assessment reproducibility was studied among 172 laboratories using tissue microarray slides with 20 tissue spots negative and 10 tissue spots expressing ER at low, medium, or high levels. More than 80% of the laboratories demonstrated ER positivity in the medium- and high-expressing tissue spots, but only about 43% succeeded with tissue spots with low expression. Poor interlaboratory agreement was based on insufficient retrieval efficacy as shown by additional tests using autoclave pretreatment. The immunohistochemical scores used to quantify therapeutic target molecules remain inconclusive as long as progress toward standardized immunohistochemical procedures and evaluation is not achieved. Tissue microarray technology has proved its suitability for large-scale immunohistochemical trials, giving rise to new dimensions in control assessment.

Female↗
PubMed2002

Detection of chromosomal imbalances in hepatocellular carcinoma.

Hepatocellular carcinoma is the most common malignant tumor of the liver. The discrimination between well-differentiated hepatocellular carcinoma and reactive lesions and benign tumors may be difficult, especially when performed on the basis of needle biopsies. A promising means of solving this problem is provided by chromosomal analysis of imbalances in hepatocellular carcinoma. This article describes the different approaches to ascertain differential diagnosis by chromosomal studies in a reliable and cost-effective manner. It is shown that in situ hybridization techniques provide a reliable means of defining chromosome alterations. These techniques allow the detection of genetic gains and losses of defined chromosomes in a histopathological context and can serve as a helpful tool in establishing diagnosis of liver cell proliferation.

Carcinoma, Hepatocellular↗
PubMedDate not supplied

Standardized on-slide control for quality assurance in the immunohistochemical assessment of therapeutic target molecules in breast cancer.

Individualization and optimization of risk-adapted therapy in breast cancer requires molecular profiling of individual cancer specimens. In tissues of heterogeneous cellular composition, such as mammary tissue, cancer prognostic markers and potential therapeutic target molecules are preferentially detected by in situ techniques such as immunohistochemistry. The more therapy decisions are based on immunohistochemical findings, the more histopathologists are confronted with the demand to establish standardized procedures that enable reproducible evaluation independent from the investigating laboratory. Disappointing results of recent national and international immunohistochemistry trials for steroid receptor and HER-2 analysis in breast cancer underline the need to act now beyond recommendations and well-formulated guidelines for standardization. In order to ensure staining quality in every individual immunohistochemical analysis, we have developed an on-slide control with standardized samples which are placed on every slide of a therapy-decisive stain together with the tissue of interest. Novel mini tissue microarrays were constructed comprising cell lines as standardized controls, indicating the sensitivity and reliability of the staining procedure. Standardized control material on the same slide as the patient's specimen require neither additional reagents nor time and are automatically archived on the diagnostic slide to prove the quality of staining even after years.

Biomarkers, Tumor↗