Papers by Ulrich-peter Rohr
BMC Health Services Research, 2015
Treatment for patients with breast cancer (BC) is guided by human epidermal growth factor recepto... more Treatment for patients with breast cancer (BC) is guided by human epidermal growth factor receptor 2 (HER2) status. The patient's HER2 status is assessed using US Food and Drug Administration-approved in vitro diagnostic (IVD) immunohistochemical (IHC) tests and laboratory-developed IVD tests. We analysed HER2 testing accuracy using data from the Nordic Immunohistochemistry Quality Control (NordiQC) HER2 IHC programme; results were used in an economic BC treatment model. Data were obtained from NordiQC HER2 BC surveys performed from 2008 to 2012. False-negative (FN) and false-positive (FP) rates for approved and laboratory-developed IVDs were used to estimate direct costs, loss of survival, productivity benefit and quality-adjusted life-years. In the absence of consistent and accessible clinical and economic data from countries participating in the NordiQC programme, United States productivity data, healthcare costs and patient numbers were used as a surrogate in order to estimate the potential impact of selecting an approved or laboratory-developed IVDs. In total, 1703 tests were performed. Pooled FN rates were 11 % for approved IVDs and 25 % for laboratory-developed IVDs; FP rates were 0 % and 5 %, respectively. Using these FP and FN rates in the economic model and applying them to the United States BC population, approved IVD tests would result in better clinical outcomes, i.e., better survival and fewer disease recurrences/progressions, and lower costs, i.e., total direct costs and lost productivity, versus laboratory-developed IVD tests. Every $1 saved by laboratories by using cheaper reagents could potentially result in approximately $6 additional costs to the healthcare system. The results of this analysis suggest that incorrect HER2 test results have far-reaching clinical and economic consequences.
Blood
CD34+ hematopoietic stem cells are used clinically to support cytotoxic therapy, and recent studi... more CD34+ hematopoietic stem cells are used clinically to support cytotoxic therapy, and recent studies raised hope that they could even serve as a cellular source for nonhematopoietic tissue engineering. Here, we examined in 18 volunteers the gene expressions of 1185 genes in highly enriched bone marrow CD34+ (BM-CD34+) or granulocyte-colony-stimulating factor-mobilized peripheral blood CD34+ (PB-CD34+) cells by means of cDNA array technology to identify molecular causes underlying the functional differences between circulating and sedentary hematopoietic stem and progenitor cells. In total, 65 genes were significantly differentially expressed. Greater cell cycle and DNA synthesis activity of BM-CD34+ than PB-CD34+ cells were reflected by the 2- to 5-fold higher expression of 9 genes involved in cell cycle progression, 11 genes regulating DNA synthesis, and cell cycle-initiating transcription factor E2F-1. Conversely, 9 other transcription factors, including the differentiation blockin...
Clinical laboratory
testingdiagnostics ] I n an era of medicine noted for the development of targeted therapies and p... more testingdiagnostics ] I n an era of medicine noted for the development of targeted therapies and personalized health care (PHC), the field of diagnostics has unprec-edented opportunities to make a valuable con-tribution to patient care. It is widely accepted today that a PHC model—enabled by high-value diag-nostic tests and molecularly targeted therapies— will be the key to driving further progress in the treatment of many diseases, particularly cancer. This recognition of the extraordinary potential PHC has to influence patient outcomes is, in turn, causing the value of diagnostic tests to increase across the entire spectrum of patient care, from screening to therapy monitoring. Rather than sim-ply providing information regarding the presence other therapeutic areas in terms of patient response rates to major drugs (Figure 1, page 10). Similarly, while cancer is comprised of many distinct disease states, the category in general has not seen signifi-cant advances in the medical value...
Deutsche medizinische Wochenschrift (1946), Jan 30, 2001
HISTORY, ADMISSION FINDINGS AND DIAGNOSIS: After stem-cell transplantation a 45-year-old woman (c... more HISTORY, ADMISSION FINDINGS AND DIAGNOSIS: After stem-cell transplantation a 45-year-old woman (case 1) had an attack of general hypoxia requiring resuscitation. She then developed a quadriplegia and spasticity of all limbs notably of the right arm and a severe pain syndrome which had to be treated by oral and intravenous analgesics. Immobilisation and secondary complications aggravated the already difficult situation. In the 2nd case a 66-year-old woman was admitted to our outpatient clinic with long-standing left-sided spastic hemiparesis after territorial infarction of the right middle cerebral artery. Beside the spasticity she also suffered from a distinct pain syndrome which did not respond to any oral analgesics. For the treatment of the main symptoms, both patients received intramuscular injections of 1000 MU botulinum toxin A (Dysport(R) Ipsen Pharma). Astonishingly, both patients experienced pain relief the next day, whereas spasticity started to respond only 5-6 days later...
Verhandlungen der Deutschen Gesellschaft für Pathologie, 1998
As many other nuclear markers, e.g. steroid receptors, Ki-67 epitopes are differentially expresse... more As many other nuclear markers, e.g. steroid receptors, Ki-67 epitopes are differentially expressed in tumour cell nuclei. It is unclear whether this phenomenon represents tumour cell heterogeneity, different stages of the cell-cycle or a biological phenomenon with prognostic impact. We analysed 104 primary adult soft tissue sarcomas (ASTS), formalin-fixed, paraffin-embedded, by APAAP and LSAB immunohistochemistry, epitope retrieval technique and 2 anti-Ki-67 antibodies (MIB-1 and Ki-S-5). Expression was evaluated by 4 indexes/1000 tumour cells: a) A-index: sum of all (weak, moderate and strong stained) Ki-67-nuclei, b) the weighed R-index: sum of all strong stained Ki-67+ nuclei x3, moderate stained nuclei x2 and weak stained nuclei x1, c) ID1-index: sum of all strong stained Ki-67+ nuclei, and d) ID2-index: sum of all strong and moderate stained Ki-67+ nuclei. Prognostic impact was analysed by Kaplan-Meier and logrank statistics with respect to overall survival. Quantitative Ki-67 ...
DMW - Deutsche Medizinische Wochenschrift, 2001
Clinical cancer research : an official journal of the American Association for Cancer Research, 2005
The fragile histidine triad protein (FHIT) is a putative tumor suppressor in patients with lung c... more The fragile histidine triad protein (FHIT) is a putative tumor suppressor in patients with lung cancer. In this study, we examined the prognostic value of FHIT expression for survival in patients with small cell lung cancer (SCLC). As assessed by immunohistochemistry using formalin-fixed, paraffin-embedded tissue sections, tumors of 225 patients with SCLC were retrospectively evaluated for FHIT expression. The influence of FHIT staining intensities as well as the proportion of FHIT-positive cells within a tumor was taken into consideration for univariate and multivariate survival analysis. FHIT expression was observed in 61.8% of the SCLC tumors. Lack of FHIT was significantly associated with a shorter survival time for the patients with a median of 157 +/- 18 days compared with 210 +/- 18 days for those patients with FHIT-positive tumors (P = 0.0061). Furthermore, the proportion of FHIT-positive cells within the tumor was related to survival. Patients with tumors of <25% FHIT-po...
Journal of carcinogenesis, 2006
PTTG-1 (pituitary tumor transforming gene) is a novel oncogene that is overexpressed in tumors, s... more PTTG-1 (pituitary tumor transforming gene) is a novel oncogene that is overexpressed in tumors, such as pituitary adenoma, breast and gastrointestinal cancers as well as in leukemia. In this study, we examined the role of PTTG-1 expression in lung cancer with regard to histological subtype, the correlation of PTTG-1 to clinical parameters and relation on patients' survival. Expression of PTTG-1 was examined immunohistochemically on formalin-fixed, paraffin-embedded tissue sections of 136 patients with small cell lung cancer (SCLC) and 91 patients with non-small cell lung cancer (NSCLC), retrospectively. The intensity of PTTG-1 expression as well as the proportion of PTTG-1 positive cells within a tumor was used for univariate and multivariate analysis. PTTG-1 expression was observed in 64% of SCLC tumors and in 97.8% of NSCLC tumors. In patients with SCLC, negative or low PTTG-1 expression was associated with a shorter mean survival time compared with patients with strong PTTG-1...
Transfusion, 2006
BACKGROUND: Current regimens for peripheral blood progenitor cell (PBPC) mobilization in patients... more BACKGROUND: Current regimens for peripheral blood progenitor cell (PBPC) mobilization in patients with multiple myeloma are based on daily subcutaneous injections of granulocyte-colony-stimulating factor (G-CSF) starting shortly after cytotoxic therapy. Recently a polyethylene glycol-conjugated G-CSF (pegfilgrastim) was introduced that has a substantially longer t 1/2 than the original formula. STUDY DESIGN AND METHODS: The use of pegfilgrastim was examined at two dose levels for PBPC mobilization in patients with Stage II or III multiple myeloma. Four days after cytotoxic therapy with cyclophosphamide (4 g/m 2 ), a single dose of either 6 mg pegfilgrastim (n = 15) or 12 mg pegfilgrastim (n = 15) or daily doses of 8 µg per kg unconjugated G-CSF (n = 15) were administered. The number of circulating CD34+ cells was determined during white blood cell (WBC) recovery, and PBPC harvesting was performed by large-volume apheresis. RESULTS: Pegfilgrastim was equally potent at 6 and 12 mg with regard to mobilization and yield of CD34+ cells. No dose dependence was observed because CD34+ cell concentration peaks were 131 and 85 per µL, respectively, and CD34+ cell yield was 10.2 × 10 6 and 7.4 × 10 6 per kg of body weight, respectively. Pegfilgrastim in either dose was associated with a more rapid WBC recovery (p = 0.03) and an earlier performance of the first apheresis procedure (p < 0.05) in comparison to unconjugated G-CSF. No difference regarding CD34+ cell maximum and yield could be observed. CONCLUSION: A single dose of 6 mg pegfilgrastim is equally potent as 12 mg for mobilization and harvest of PBPCs in patients with multiple myeloma. Because no dose dependency was seen at these dose levels, this might be also true for even smaller doses.
Journal of Virological Methods, 2005
In this report, we present a fast, reliable and easy to perform method to quantify infectious tit... more In this report, we present a fast, reliable and easy to perform method to quantify infectious titers of recombinant AAV-2 (rAAV-2) particles using the LightCycler technology, which is independent from the therapeutic transgene and without the presence of a marker gene. The method is based on the life cycle of AAV-2: after infection of the host cell, the single stranded (ss) AAV-2 genome is converted into a double stranded (ds) form. Following infection with rAAV-2, HeLa cells were lysed and ssDNA of transcriptionally inactive particles were efficiently removed by ssDNA-specific S1 nuclease digestion. The remaining viral dsDNA can be quantified by quantitative real-time PCR (qPCR). For validation of the new method, rAAV-2 preparations were analyzed by two other standard methods for titration of infectious particles in parallel, i.e. the infectious center assay (ICA) as well as flow cytometry using GFP as a marker. Comparing the infectious titers of 40 different AAV-2 fractions assessed by qPCR with the titers determined by FACS analysis a significant correlation (r=0.87, p&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt;0.001) with a mean ratio of the titers assessed by qPCR and FACS of 1.92 (S.D.+/-1.59) was found. Further, the titers of seven rAAV-2 fractions using qPCR and ICA covering 5 log ranges were compared and a significant correlation was found between the results (r=0.80, p&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt;0.001) with a mean ratio of 3.38 (S.D.+/-1.79), respectively.
Journal of Virological Methods, 2002
The susceptibility of a variety of different primary tissues was examined to long-term transducti... more The susceptibility of a variety of different primary tissues was examined to long-term transduction with recombinant adeno-associated virus type 2 (rAAV-2) and factors influencing the transduction efficiency. In contrast to others using cell lines and animal models, emphasis was placed on the use of primary human cells. Enhanced green fluorescent protein (EGFP) marker gene expression was examined using fluorescence-activated cell sorting analysis. The most effective target cells for rAAV-2-mediated gene transfer were bronchial epithelial, artery endothelial as well as smooth and skeletal muscle cells with mean transduction rates ranging from 34.3 to 81.6%. Lower transduction rates between 4.3 and 19.5% were found in chondrocytes, dermal papilla follicle epithelial cells and fibroblasts. No transduction was observed in melanocytes, granulocyte colony-stimulating factor (G-CSF)-mobilized CD34(+) cells or malignant CD19(+) cells from patients with chronic lymphocytic leukemia. A proportion of EGFP-expressing skeletal muscle and smooth muscle cells was maintained over a period of 6 weeks after transduction (42.7+/-5.4 and 67.1+/-0.9%, respectively). Interestingly, among hair follicle epithelial cells the proportion of transduced cells increased from 8+/-0.5 to 36+/-7.7% in the course of 6 weeks. In contrast, for endothelial cells, bronchial epithelial cells and fibroblasts, a rapid decline in the number of EGFP expressing cells were noted. An inverse relationship between the proportion of cells in G2/M phase of cell cycle and long-term gene expression was observed. All rAAV-2 susceptible primary cells expressed FGFR-1 and the alphaV integrin consistent with their role as co-receptors for AAV-2. In conclusion, AAV-2 is a suitable vector system for transduction and evaluation of functional effects of long-term gene expression in primary human muscle and hair follicle cells.
Journal of Virological Methods, 2002
In this study, a quantitative real-time PCR (qPCR) was developed to determine genomic rAAV-2 tite... more In this study, a quantitative real-time PCR (qPCR) was developed to determine genomic rAAV-2 titers using the Light-Cycler technology. Since the CMV promoter is the most commonly used promoter in gene therapeutic approaches, primers were designed which hybridize with the human CMV promoter sequence. PCR products were detected by the addition of SYBR green. qPCR of a 5 log spanning serial dilution of the vector plasmid containing one CMV promoter per plasmid molecule yielded a high amplification efficiency of 1.99 per cycle. To quantify the copy number of viral genomes, the qPCR curves of adeno-associated virus type 2 (AAV-2) samples were related to a standard curve assessed by the 5 log spanning serial vector plasmid dilution (0.01-100 pg DNA). For validation of the method, rAAV-2 preparations were analyzed by a standard method and qPCR in parallel. As standard method, flow cytometry was used for titration of infectious viral particles on HeLa cells using the Enhanced Green Fluorescent Protein as a marker. A significant correlation was found between the results obtained by flow cytometry and the results from the qPCR over a 5 log range (r=0.85, P&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt;0.0001). The mean ratio between infectious rAAV-2 particles titrated via flow cytometry and genomic copies of rAAV-2 measured by qPCR of the same sample was 1:253. The higher titers found by qPCR might be due to multiple transduction of a single cell or to non-infectious particles generated during rAAV-2 preparation. In conclusion, qPCR is a fast and reliable method for determination of rAAV-2 titers and might be a powerful tool for standardization of rAAV-2 preparations particularly in the context of clinical studies.
Journal of Translational Medicine, 2008
We examined gene expression profiles of tumor cells from 29 untreated patients with lung cancer (... more We examined gene expression profiles of tumor cells from 29 untreated patients with lung cancer (10 adenocarcinomas (AC), 10 squamous cell carcinomas (SCC), and 9 small cell lung cancer (SCLC)) in comparison to 5 samples of normal lung tissue (NT). The European and American methodological quality guidelines for microarray experiments were followed, including the stipulated use of laser capture microdissection for separation and purification of the lung cancer tumor cells from surrounding tissue.
Journal of Clinical Oncology, 2011
TO THE EDITOR: We read the article by Hurwitz et al 1 on the venous thromboembolic events (VTEs) ... more TO THE EDITOR: We read the article by Hurwitz et al 1 on the venous thromboembolic events (VTEs) with chemotherapy plus bevacizumab in patients with cancer that was based on a meta-analysis of individual patient data with great interest. We would like to make three comments that are relevant to the conclusion of the study, which is not consistent with our previous analysis. First, the overall conclusion of bevacizumab's effect on VTEs depends on the studies included for the meta-analysis. This study included 10 randomized controlled clinical trials, which may be a small fraction of total bevacizumab trials. The article did not mention how many studies the authors had screened before the final selection of these trials. The difference in the number of included trials between this study and our previous meta-analysis 2 is substantial; although the two meta-analyses shared the same eight studies, they differed in a total of nine trials; the current study had added two new studies by Miles et al 3 and Van Cutsem et al 4 with a calculated summary risk ratio of 0.82 (95% CI, 0.60 to 1.11) and did not have seven other trials with asummaryrelativeriskof1.27(95%CI,0.94to1.73)whichwereincluded in our study. 2 Thus, the determination of overall RR by this study will be most likely underestimated by the addition and exclusion of these studies.
Journal of Cancer Research and Clinical Oncology, 2010
Background Bevacizumab is frequently combined with 5-Xuorouracil-based chemotherapy for patients ... more Background Bevacizumab is frequently combined with 5-Xuorouracil-based chemotherapy for patients with metastatic colorectal cancer (mCRC). The relative beneWt of bevacizumab in older patients has not been widely studied and is of interest. Patients and methods This retrospective analysis used data from three Wrst-line randomized controlled studies and one second-line randomized controlled study of bevacizumab plus chemotherapy in medically Wt (Eastern Cooperative Oncology Group performance status 0 or 1) patients with mCRC. Overall survival (OS) and on-treatment progression-free survival (PFS) were assessed in patients aged <65, ¸65, and ¸70 years. Results were compared using unstratiWed hazard ratios (HRs). Grade 3-5 adverse events were also assessed.
International Journal of Cancer, 2005
Aberrant promoter methylation of normally unmethylated CpGislands offers a promising tool for the... more Aberrant promoter methylation of normally unmethylated CpGislands offers a promising tool for the development of molecular biomarkers. We investigated bronchial aspirates of patients admitted for suspected lung cancer with regard to the prevalence of aberrant methylation of potential marker genes. Applying quantitative methylation specific PCR (QMSP) we analyzed bronchial aspirates from 75 patients with primary lung cancer and 64 bronchial aspirates of patients diagnosed with benign lung disease for promoter methylation of 3 candidate marker genes (p16 INK4a , RARB2 and SEMA3B). Hypermethylation of p16 INK4a detected 18/75 (24%) cases with primary lung cancer and was present predominantly in squamous cell carcinomas (14/25; 56%). RARB2 QMSP at an assay threshold greater than 30 was found in 42/75 (56%) patients with lung cancer without relation to histological subtype. Patients with benign lung disease showed methylation of p16 INK4a and a RARB2 QMSP at an assay threshold greater than 30 in 0/64 (0%) and 8/64 (13%) cases, respectively. Combining the 2 methylation markers, p16 INK4a and RARB2, yielded a sensitivity of 69% and a specificity of 87% for the diagnosis of pulmonary malignancy. In contrast, SEMA3B displayed frequent promoter methylation (around 90%) both in bronchial aspirates of tumor and nontumor cases and thus was not suited as a biomarker. The results of this study indicate that QMSP analysis of p16 INK4a and RARB2 may aid the diagnosis of primary lung cancer in bronchial aspirates. In particular, detection of p16 INK4a methylation by QMSP may serve as a highly specific marker of pulmonary squamous cell carcinoma. ' 2005 Wiley-Liss, Inc.
International Journal of Cancer, 2004
In a retrospective analysis of 203 patients with small cell lung cancer (SCLC), we examined the p... more In a retrospective analysis of 203 patients with small cell lung cancer (SCLC), we examined the prognostic value of c-kit expression on survival. Expression of c-kit was examined immunohistochemically in formalin-fixed, paraffin-embedded tissue sections. c-kit was observed in 87.7% of SCLC tumors. Using the Kaplan-Meier model, we found that lack of c-kit expression was associated with significantly shorter survival time compared to the presence of c-kit expression (mean survival 151 ؎ 27 vs. 358 ؎ 49 days, p ؍ 0.0084). Moreover, the proportion of c-kit ؉ cells within the tumor was also related to survival time. Patients with tumors in which >75% of cells stained positive for c-kit had a mean overall survival time of 424 (؎72) compared to 295 (؎67) days for patients with 25-75% c-kit ؉ tumor cells. Patients with tumors containing <25% c-kit ؉ cells had the worst survival, with 164 (؎24) days (p ؍ 0.0033). Further parameters associated with short survival times were low performance status, elevated levels of lactate dehydrogenase and higher stage according to the TNM classification. Multivariate analysis using the Cox regression model showed that the proportion of c-kit ؉ cells within the tumor specimen was one of 3 independent prognostic parameters (p ؍ 0.004) for overall survival next to TNM classification (p ؍ 0.001) and performance status (p < 0.001).
Drug Discovery Today: Therapeutic Strategies, 2013
Clinical Endocrinology, 2006
Objective Adrenocortical carcinoma (ACC) is a rare malignancy associated with a dismal prognosis.... more Objective Adrenocortical carcinoma (ACC) is a rare malignancy associated with a dismal prognosis. Dendritic cells (DCs) are professional antigen-presenting cells leading to an antitumour immune response. The aim of this study was to elaborate two methods of antigen delivery to DCs and to evaluate an immunotherapy protocol in ACC patients. Design/patients Autologous DCs were pulsed with autologous tumour lysate (TL). Fusion of DCs with tumour cells was based on a polyethylene glycol method. Two patients with metastasized hypersecretory ACC were vaccinated twice. Measurements In vitro data were quantified by measurement of PBMC (peripheral blood mononuclear cell) responses and cytokine secretion and by flow cytometry analyses. Clinical response was monitored by CT scan of tumour mass and measurement of angiogenic factors. Results The maximum loading of TL was obtained at 24 h as 48·2% ( ± 26·8%) of DCs were TL-positive. The DC/tumour cell fusion efficacy was ∼ 45% as shown by double positive staining for ACTH receptor and DC-specific CD83. In vivo DC vaccination resulted in positive delayed-type hypersensitivity skin reactions reflecting specific memory T-lymphocyte reaction. In vitro analyses revealed specific T-cell proliferation in patient 1 (stimulation index: 5·7 compared to pretreatment) and induction of cytotoxic granzyme B secreting T cells in patient 2 (0·41% CD8 + cells vs. 0·06% pretreatment) as indicators of specific cytotoxic T cells. Although angiogenic serum markers could be stabilized, no impact on tumour growth could be observed. Conclusion Our data demonstrate that autologous dendritic cells induce antigen-specific Th1 immunity in adrenocortical carcinoma. The clinical outcome, however, was not improved in the patients studied here.
Clinical Cancer Research, 2010
Patients with central nervous system (CNS) metastases were excluded from bevacizumab trials follo... more Patients with central nervous system (CNS) metastases were excluded from bevacizumab trials following a case of fatal cerebral hemorrhage in a patient with hepatocellular carcinoma in 1997. Safety information for bevacizumab-treated patients with CNS metastases was reviewed to determine whether general exclusion of these patients from bevacizumab treatment is still justified. A retrospective exploratory analysis was conducted using datasets from 13 randomized controlled phase II/III trials (dataset A), two open-label single-arm safety trials (dataset B), and two prospective studies including patients with treated CNS metastases (dataset C). In datasets A and B, known CNS metastasis was an exclusion criterion; patients with CNS metastasis had unrecognized CNS metastases at study entry or developed them during the trial. All reported cerebral hemorrhage grades in patients with CNS metastases were quantified. In dataset A, occult brain metastases were identified in 187 of 8,443 patients (91 in bevacizumab arms and 96 in non-bevacizumab arms). Three bevacizumab-treated patients (3.3%) developed grade 4 cerebral hemorrhage, whereas one control-arm patient (1.0%) developed grade 5 cerebral hemorrhage. In dataset B, 321 of 4,382 patients had initially occult CNS metastases, in whom two grade 1 and one grade 3 cerebral hemorrhage (0.9%) were reported. In 131 patients with treated CNS metastases in dataset C, one bevacizumab-treated patient (0.8%) developed grade 2 cerebral hemorrhage. In this selected population, patients with CNS metastases are at similar risk of developing cerebral hemorrhage, independent of bevacizumab therapy. Consequently, such patients with CNS metastases from advanced/metastatic breast cancer, non-small cell lung carcinoma, and renal and colorectal cancer should not be generally excluded from bevacizumab therapy or clinical trials.
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Papers by Ulrich-peter Rohr