bluuebunny/crossref_metadata_2025 huggingface dataset:
Prognostic significance of immunohistochemical expression of Rb gene product in operable breast cancer
Authors: Song H. , Do Y. , Gang S. , Kwon S. , Kim S. , Lee W. , Lee J.
Published: March 2020

21121

Background: The aim of this study was to investigate the prognostic significance of the expression of Rb gene product in operable invasive breast cancer by performing immunohistochemical analysis. Methods: Between January 1993 and December 2001, 212 operable invasive breast cancer patients underwent immunohistochemical staining for Rb, and we retrospectively analyzed these results together with the clinical outcomes. Results: The overexpression of p53 was detected in 72.7% of the cases. The overexpression of Rb was correlated with positive hormonal receptor (p=0.000), and inversely correlated with lymph node metastasis (p=0.017) and vascular invasion (p=0.004). The tumor size, tumor histology, histologic grade, and tumor stage were not related to the overexpression of p53. Multivariate Cox regression analysis indicate that lymph node metastasis and tumor size were the significant prognostic factors for overall survival; lymph node metastasis was the significant prognostic factor for relapse free survival. On the subgroup analysis, the Rb expressors showed better 7-year overall survival (98.5% vs. 81.5%, respectively, p=0.005) and relapse free survival (94.1% vs. 77.4%, respectively, p=0.021) than did the p53 non-overexpressors for the patients without lymph node metastasis. However, for the patients with lymph node metastasis, the survival rates were not different for both the Rb expressors and the Rb non-expressors. Conclusions: Immunohistochemical staining of the Rb gene product was an independent prognostic factor for predicting survival of the lymph node negative operable breast cancer patients.

No significant financial relationships to disclose.
Expression of matrix metalloproteinases (MMP1, 2, 9, and 11) in metastatic lymph node tissue of breast cancer patients
Authors: Fink-Retter A. , Gschwantler-Kaulich D. , Hudelist G. , Walter I. , Czerwenka K. , Singer C.
Published: March 2020

21138

Background: Invasive growth requires degradation of extracellular matrix. A number of Zn 2+ -dependent matrix metalloproteinases (MMPs) is associated with this process and leads to local invasion and metastasis in malignant breast tumors. Especially the expression of MMP-2 and MMP-9 has been associated with high potential of metastasis in breast cancer. Methods: We determined the expression patterns of epithelial (E) and stromal (S) MMP-1, MMP-2, MMP-9 and MMP-11 by immunohistochemistry in tissue arrays, containing 50 paraffin-embedded sets of tissues obtained from the malignant tumor of the breast, and 50 paraffin-embedded sets of tissues from metastatic lymph nodes (purchased from Biomax, Biomax Inc, Rockville, MD). Results: MMP2-S and MMP9-S protein expression was significantly higher in lymph node tumor tissue when compared to breast tumor tissue (10% vs. 2%; p=0.005; 78.3% vs. 94.8%, p=0.01 Chi Square test, respectively). MMP9-E showed a borderline significance in lymph node tumor tissue (92.6% vs. 73.5%; p=0.05 Chi Square test). While in breast tumor tissue significant correlations were observed between all MMPs except MMP2-S, these associations were less pronounced in lymph node tumor tissue. Conclusions: We suggest that the expression of MMP-2 and MMP-9 in breast cancer tissue is associated with a high invasive and metastatic potential. This might be confirmed by our finding that MMP2-S, MMP9-S and MMP9-E are increased expressed in metastatic lymph node tissue. This is in line with other studies which showed that MMP-2 and MMP-9 expression in primary breast cancer tissue samples was associated with breast cancer progression and invasion.

No significant financial relationships to disclose.
Semi-quantitative gene expression profiling for therapy prediction in a breast cancer neoadjuvant therapy study applying docetaxel/epirubicin/cyclophosphamide
Authors: Vogt U. , Kemming D. , Brandt B. , Bosse U. , Bonk U. , Assen A. v. d. , Muhs H. J. , Schlotter C. M.
Published: March 2020

21144

Background: More than 28000 curated human genes can be analyzed semi-quantitatively using a chemiluminescent detection technology and 60mer oligonucleotides on a Human Genome Survey Microarray (HGSM, Applied Biosytems). Methods: HGSM expression profiling was performed on biopsy samplesfrom a setting of patients under neoadjuvant treatment. The protocol for a phase II study was elaborated for the treatment of breast cancer patients suffering from a primary tumor > 1,5 cm or inflammatory breast cancer with Docetaxel / Epirubicin/Cyclophosphamide (TEC) prior to surgical treatment. The study was approved by the local ethical committee and 80 patients will be included into the study after written informed consent. 40 patients have been already included in the ongoing study. In this prospective study a biopsy from the tumor is taken before chemotherapy. Therefore, success of treatment is detectable directly at the operated residual tumor. Our results demonstrate a low rate of false-positives (1.2%), a high specificity and quantification accuracy of HGSM system. Comparison of data from HGSM and RT-PCR obtained on mRNA from fresh frozen tissue resulted a Pearson-correlation of 0.92 to 0.63 for the breast cancer genes. Results: Tumour response (pCR, pPR) of more than 70 % can be achieved using neoadjuvant TEC- regimen. 25 % of pCR in this study is comparable with data of other published neoadjuvant trails. First expression profiling results are obtainable showing that a subset of 148 genes indicates patients with complete remission (pCR, no detectable tumor at end of chemotherapy), partial remission (pPR) and progressive disease (pPD). Remarkable, that the expression profile clearly separated pCR and pPD tumors whereas pPR tumors presented with a closer relationship to pPD tumors than to pCR tumors but displayed small specific subprofiles. A comparable separation of the groups could not achieved by established tumor markers, e.g. ER, PgR, HER2, uPA etc. which are measured simultaneously on the HGSM. Conclusions: HGSM expression profiling is promising to have the potential to figure out genes that are related to cancer progression and chemotherapy resistance, especially in PST.

No significant financial relationships to disclose.
Clinicopathological features in correlation with epidermal growth factor receptor gene mutations in non-small cell lung cancer (NSCLC)
Authors: Tanaka F. , Matsumoto S. , Takuwa T. , Kondo N. , Okumura Y. , Fukuoka K. , Nakano T. , Hasegawa S.
Published: March 2020

21148

Background: Mutations in the epidermal growth factor receptor (EGFR) gene are found in 10–30% of non-small cell lung cancer, and are useful to predict the efficacy of EGFR-tyrosine kinase inhibitors (EGFR-TKIs) such as gefitinib and erlotinib. However, it is sometimes difficult to examine such mutations. Thus, prediction of presence or absence of EGFR mutations, correlated with clinical response to EGFR-TKIs, based on clinicopathological characteristics may be important in clinical practice. Methods: We retrospectively assessed EGFR-mutation status in correlation with clinical features including FDG-PET findings and pathological features including presence or absence of bronchioalveolar component (BAC) in adenocarcinoma (Ad) case. A total of 72 consecutive patients (24 females; mean age, 65 years; 40 Ad cases) who received thoracotomy from August 2005 through October 2006 were reviewed Results: Sixteen cases (22%) had EGFR-mutations (8 cases each in exon 19 and exon 21). The incidence of EGFR-mutation was significantly higher in female (42%) or never-smoker (60%) than in male (13%) or smoker (15%). EGFR-mutation was almost exclusively found in Ad; among Ad cases, there was no significant difference in the incidence of EGFR-mutation between BAC-positive (47%) and BAC-negative cases (29%; p=0.33). Among 61 cases who received FDG-PET scan prior to thoracotomy, only 15% of EGFR- mutated cases had higher FDG-uptake, whereas 47% of EGFR-wild cases had higher FDG-uptake (p=0.36); the sensitivity and specificity of lower FDG-uptake to predict the presence of EGFR-mutations were 71% and 53%, respectively. The positive predictive value (PPV) of lower- FDG-uptake for the presence of EGFR-mutations was only 29%, but the negative predictive value (NPV) was as high as 85%. The mean standardized uptake value (SUV) of FDG for EGFR-wild patients and EGFR-mutated patients were 5.7 and 3.3, respectively (p=0.11). The mean SUV for tumors mutated in exon 19 seemed to be lower than that for tumors mutated in exon 21 (1.8 versus4.6). Conclusions: Higher FDG-uptake combined with clinical features (male and smoker) may help to exclude patients with EGFR-wild tumors who may not respond to EGFR-TKIs treatment.

No significant financial relationships to disclose.
Molecular markers to differentiate between pancreatic cancer and normal pancreas/chronic pancreatitis: Gene expression profiling study
Authors: Jarzab M. , Olakowski M. , Oczko-Wojciechowska M. , Kowalska M. , Fujarewicz K. , Chmielik E. , Lange D. , Świerniak A. , Jarz'ab B. , Lampe P.
Published: March 2020

21150

Background: The aim of the study was to analyze the gene expression profile of pancreatic cancer by multivariate methods of class prediction. Methods: The snap-frozen or RNA-later preserved samples of 18 pancreatic adenocarcinomas, 9 chronic pancreatitis cases and 6 specimens collected from microscopically unchanged pancreas (N/CP) were analyzed by HG-U133 Plus 2.0 oligonucleotide microarrays (Affymetrix). The obtained dataset was pre-processed using GC-RMA method, gene selection was carried out both by class comparison methods (Welch test with Benjamini-Hochberg correction, False Discovery Rate FDR<5%) and by our own algorithms of class prediction, based on Support Vector Machines technique (Recurrent Feature Replacement and Bootstrap-Based Feature Ranking). Real- time quantitative PCR (Q-PCR) was carried out on Applied Biosystems 7900 HT machine, with Universal Probe Library (Roche) fluorescent probes and normalization by three reference genes index (geNorm, Vandesompele et al.). Results: We compared gene expression profiles between pancreatic cancer samples and N/CP specimens. 23850 probesets significantly differentiated between these three classes (FDR<5%). No ideal discrimination between cancer and N/CP samples was possible by any of single markers. We selected the optimal multi- gene classifier by Support Vector Machines, using Bootstrap-Based Feature Ranking method. The smallest classifier resulting in 100% accuracy consisted of three genes, 45 genes were included in more than half of the diagnostic genesets obtained during bootstrapping process. 14 genes were selected for Q-PCR validation, again none of them ideally discriminated between cancer and normal specimens, with the area under the receiver-operating-characteristic curve ranging from 0.82–0.93. Three-gene combinations allowed for proper classification of all samples. Conclusions: The multi-gene classifier, derived both by microarray technique and Q-PCR analysis, is properly discriminating between pancreatic cancer and chronic pancreatitis/normal pancreas. At least three genes must be included in the classifier to obtain satisfying accuracy. MO and MJ equally contributed to the study.

No significant financial relationships to disclose.
Epitope mapping of epidermal growth factor receptor (EGFR) in lung cancer using immunohistochemistry: Fine tuning the protocols
Authors: TIMAR J. , Derecskei K. , Dome B. , Moldvay J.
Published: March 2020

21162

Background: Until now, immunohistochemistry was not able to become a reliable diagnostic approach for EGFR targeted therapies. The golden standard of the determination of EGFR protein expression in paraffin embedded cancer tissues is the EGFRpharmDXtm kit. Methods: Here we show data based on analysis of 110 lung adenocarcinomas, that the recommended protocol may not be optimal for ideal performance of the immunodetection, since microwave retrieval, extended primary antibody-incubation time and replacement of the developer reagent converted four EGFR-negative tumor into EGFR protein positive out of eight lung adenocarcinoma cases. Protocol modification improved the performance of another widely used EGFR-kit, Ventana's CONFIRM, where replacement of the protease antigen retrieval with microwave cooking converted several EGFR-negative tumors to strongly positive. Meanwhile both EGFR-kits detect EGFR expression (juxtamembrane domain) but do not provide information on the expression of epitopes critical from the point of view of targeted therapy. Results: We have developed two protocols, which can detect the ligand-binding (AB-10, BioMarkers) and C-terminal (AB- 335, Biogenex) cytoplasmic domains of the EGFR protein in paraffin embedded lung cancer tissues. We have shown, based on the analysis of more than 110 lung adenocarcinoma tissues, that the ligand binding domain of EGFR is rarely expressed while the C-terminal domain is ubiquitously expressed in EGFR-PharmDX and CONFIRM-positive cancers. The biological activity of EGFR can be characterized either by autophosphorylation of the receptor or by detection of divers phosphorylated downstream signaling components. We have found that unlike p1086 (detected by a Zymed antibody), the p1173 site of EGFR (identified by a rabbit monoclonal of Epitomics) can be detected 27/110 paraffin embedded lung adenocarcinomas. Conclusions: Using the tested antibody panel we can reliably determine the EGFR protein expression in paraffin embedded (lung)cancer tissues. This work was supported by Ministry of Education (NKFP1a-0024–05).

No significant financial relationships to disclose.
The alternation of estrogen receptor (ER) and progesterone receptor (PgR) expression in primary breast cancer patients treated with neoadjuvant chemotherapy (NAC)
Authors: Kitagawa D.
Published: March 2020

21165

Background: Currently, neoadjuvant chemotherapy (NAC) for the operable primary breast cancer has become standard treatment strategy. Primary treatment could affect the status of Estrogen receptor (ER) and Progesterone receptor (PgR) expressions in the tumor, and might have impact on the choice of adjuvant hormone therapy. Methods: We evaluated consecutive 165 primary breast cancer cases those treated with 4 cycles of FEC (500–100–500 mg/m2, q3wks) followed by 4 cycles docetaxel (75 mg/m2, q3wks) as their neoadjuvant treatment in our institution from January 2000 to February 2006. Except for the bilateral breast cancer cases and patients achieved pCR, 107 pairs of core needle biopsy before treatment and tumor block after surgery were evaluated by immunohistochemistry (IHC) for receptor status and analyzed by Allred scoring. Score 4 or more was designated as positive. Results: Median age of patients was 51 (23–71) years old. Hormone receptor (HR) phenotypes before treatment were following; ER+/PgR+:62(57.9%), +/ -:23(21.5%), -/ +:2(1.9%), -/ -:20(18.7%). As the HR status, 94.4 % of patients did not show any alternation for their HR evaluation. Only 4.7% of patients experienced changes from HR+ to HR-, and HR- to HR+ were extremely rare (0.9%). Although ER positivity was not changed in most of cases (95.1%), conversion from PgR + to - were observed in 35.5% of ER+/PgR+ cases. Particularly, the rate of negative conversion of PgR were found more frequently in patients with age under 50 (51.7% in <50y.o. vs. 22.6% in =50 y.o., p=0.03). Whereas, in older patients, negative conversion of PgR correlated to favorable pathological response. Conclusions: In most of patients, neoadjuvant chemotherapy did not affect the evaluation of HR status. Alternation of PgR expression seems to reflect the chemotherapy induced amenorrhea in younger patients. In the older patients, negative conversion of PgR may have some impact on pathological response.

No significant financial relationships to disclose.
A novel case of Silver Russell Syndrome (SRS) and male breast cancer
Authors: Hamm C. M. , Jagiello L.
Published: March 2020

21178

Introduction: Silver Russell Syndome (SRS) is a heterogeneous disorder of asymmetric growth retardation and typical dysmorphic features often associated with maternal disomy 7 and chromosome 17q23–24. The BRCA1 gene is found on 17q21. The relationship between SRS and breast cancer has not been described to date. Case Presentation: We report a case of a 37 year old male diagnosed with metastatic breast cancer involving the liver and bone. Patient presented with a left proximal hip fracture related to metastatic bone disease. Biopsy of the right breast lesion revealed a moderately differentiated adenocarcinoma consistent with breast primary. Abdominal ultrasound and full body bone scan revealed extensive liver and bone metastases. There was no significant family history of cancer, specifically breast nor ovarian cancer. Discussion: The BRCA2 gene is known to be responsible for DNA repair. The SRS cases associated with chromosome 17q24–25 abnormalities are known to be associated with growth hormone deletions. Further investigations into this relationship are warranted.

No significant financial relationships to disclose.
Identification of a new cancer/testis antigen from small cell lung cancer
Authors: Lee M. , Kim K.
Published: March 2020

21182

Background: Serological analysis of recombinant cDNA expression libraires has led to identification of tumor antigens, which are considered promising candidates for early detection and immunotherapy. They are recognized by the immune system of cancer patients including small cell lung cancer (SCLC) patients. Methods: SEREX method was used to screen normal testis and NCI-H889 cell line cDNA library with serum obtained from a SCLC patient who has survived over 50 months. Also SADA test was done to SCLC patients and normal serum. Results: Immunoscreening of libraries with the selected serum led to the isolation of 58 (SCLC-1 through SCLC-58) and 18 (SCLCT-1 and SCLCT-18) independent antigens from NCI-H889 and testis respectively. Of these SCLCT-1 mRNA was expressed strongly only in normal testis. According to the SADA test, SCLCT-1 was only found in SCLC patients’ sera (34%). Conclusions: On the basis of its immunogenecity in SCLC patients’ sera, and its restricted mRNA expression profile, SCLCT-1 can be considered a novel CT antigen.

No significant financial relationships to disclose.
Prospective study on prognostic significance of DNA ploidy and Ki-67 expression in colorectal cancer
Authors: Saggia C. , Santagostino A. , Forti G. , Biaggi G. , Angeli G. , Dacorsi M. , Cerrato D. , Loi G. , Porcile G. , Alabiso O.
Published: March 2020

21184

Background: The prognostic value of flow cytometry DNA ploidy and Ki-67 expression in colorectal carcinomas has not been defined yet. The study tries to correlate tumoral DNA ploidy and proliferative activity (Ki-67) with therapy response, Overall Survival (OS), Disesase Specific Survival (DSS) and Disease Free Survival (DFS). Methods: From 01/09/02 to 30/06/05, 3 samples of cancer tissue and 1 sample of normal mucosa has been collected from all operating pieces of colorectal cancer. These samples were frosen and later disgregated, treated and coloured with Propidium Iodide. DNA ploidy was evaluated by FACSCalibur cytofluorimetry. Normal mucosa tissue was our internal control. Ki-67 was evaluated by immuno-histochemistry in all tumoral samples. All the patients were cured with chemo- and/or radiotherapy in our divisions. Results: 67 patients (M/F 35/32); median age was 70; staging: 19% I, 33% II, 30% III, 18% IV. We found aneuploidy in 65,7% of carcinoma and Ki-67 median expression was 55%. DNA tumoral heterogeneity was present in 27% of patients. DNA aneuploidy correlates with advanced disease stage at diagnosis (p<0,01), with high number of metastatic lymphnodes (p<0,005) and with serological markers positivity (p<0,04). After surgery and chemotherapy 35% of the patients with aneuploid carcinoma and high proliferative activity (Ki-67>55%) do not show evident disease versus 100% of patients with DNA diploidy and lower Ki-67. Tumoral aneuploidy correlates in a significative way with lower OS (48% vs 89% of diploid patients), lower DSS (tumor death happened just in patients with aneuploid DNA in every disease stages), with lower DFS (18% vs 86% of diploid patients). Univariated analysis stated that aneuploidy determinates an Odd Ratio=5,7 to develop disease progression (p=0,033). At the moment Ki-67 expression with 55% cut-off does not seem to correlate with OS, DSS and DFS. Conclusions: Preliminar results (the study is still in progress) seem to suggest that cytofluorimetric DNA-ploidy has a prognostic and predictive significance in colo-rectal carcinomas. Ki-67 expression (immuno-histochemistry) has an uncertain significance. The small number of patients and the short follow-up do not allow us to reach any definitive conclusions, but the study is worth to go on.

No significant financial relationships to disclose.
Reproductive risk factors of breast cancer in elderly population. A single centre study
Authors: Torregrosa D. , Gironés R. , López P. , Díaz Beveridge R. , Maestu I.
Published: March 2020

21187

Background: Relationship between reproductive variables and cancer risk has been yet established. Protective effect of late age at menarche (≥ 11) and high parity (≥ 4) and the increased risk of nulliparous women, older age at first birth (AFB) (≥ 28) and older age at menopause (≥ 52) seems to act differently in pre vs. postmenopausal elderly women. The aim of this study was to describe recognized reproductive risk factors for breast cancer in a selected population of a single centre. Methods: A cross- sectional study of women diagnosed ≥ 70 years. We selected women with removed early breast cancer and on follow-up. No disease recurrences were allowed. Between January 2005 to June 2006. Described reproductive risk factors for breast cancer were collected. Tumour characteristics also. Results: 91 patients were recruited. Mean age at time of diagnosis: 76 (70–92); mean age at study 80 (71- 95). Reproductive factors: late age at menarche (12; 9–16; percentile 75: 14) and late age at menopause: 48 (29–56); 75% ≥ 52 years old were common. Nulliparous women were similar at described in other series (10%); parity 2 (0–9); percentile 75: 3;late AFB: 26 (19–40); percentile 75: 29 years. No one had taken hormonal treatment. Only 16% had familiar history of breast cancer and 9% personal history. Almost all (84%) had positive tumours for the oestrogen receptor and 76% for progesterone receptor. 91% had ductal histology and 54% had II stage. Conclusions: Discussion: unexpected associations between classical reproductive risk factors and breast cancer on elderly patients have been described in other series. It seems that late age at menarche and late age at menopause is common in elderly breast cancer population, as in our serie. Nulliparous elderly continue to have risk but seems that the protective effect of parity dissipated in these population. Older age at first birth (≥28) is consistently associated with increased breast cancer risk. Conclusion: our study is based on a high selected population and clearly has a selection bias. Our results cannot be generalized to other populations but describes our patients. The results therefore should be internally valid. And it seems to coincide with other studies.

No significant financial relationships to disclose.
Genetic variants in human carbonyl reductase 3 (CBR3) and their influence on doxorubicin pharmacokinetics in Asian breast cancer patients
Authors: Fan L. , Guo J. Y. , Wong C. I. , Lim R. , Yap H. L. , Khoo Y. M. , Iau P. , Goh B. C. , Lee H. S. , Lee S. C.
Published: March 2020

2505

Background: Human carbonyl reductase 3 (CBR3) is one of the main metabolizing enzymes to extensively reduce doxorubicin to its major active metabolite, doxorubicinol in normal and tumor tissues. Recently, the CBR3 958G>A (V244M) genetic variant has been described to alter function in vitro. We postulate that CBR3 genetic variants could contribute to the inter-individual variability of doxorubicin pharmacokinetics in breast cancer patients. Methods: We studied 101 female breast cancer patients (66 Chinese, 26 Malay, 7 Indian and 2 of other ethnic origins) who were treated with doxorubicin at 75mg/m2 every 3 weeks. Comprehensive sequencing of the 3 exons of CBR3, including the splice-site junctions was performed. Plasma concentrations of doxorubicin and doxorubicinol were analyzed during the first doxorubicin cycle. Results: Five CBR3 coding region variants (239G>A, 483C>T, 507C>T, 598G>A and 958G>A) were detected, of which 239G>A, 598G>A and 958G>A were non-synonymous. 598G>A was novel, and was found in 1 Malay patient who was heterozygous. The genotype distributions of 239G>A and 958G>A were 36%/30%/34%, and 40%/36%/24% respectively for GG/AG/AA. The 239GG variant was associated with significantly higher AUC of doxorubicinol and AUC ratio of doxorubicinol to doxorubicin than the AG and AA variants (AUC of doxorubicinol 2.18±1.37ug/ml*h (GG) vs 2.04±2.11ug/ml*h (AG), p=0.05, and 1.55±0.61ug/ml*h (AA), p=0.004; AUC ratio of doxorubicinol to doxorubicin 1.90±1.29 (GG) vs 1.72±1.34 (AG), p=0.025, and 1.45±0.67 (AA), p=0.006). Patients with the 958AA variant had significantly higher AUC of doxorubicinol than those with the 958GG variant (2.29±1.60ug/ml*h vs 1.56±0.60ug/ml*h, p=0.009). The 239GG variant was more common in our population than in Caucasians (36% vs 20%. p=0.027), while the 958AA variant was more common than reported in Caucasians (24% vs 8%, p=0.014) and Japanese (24% vs 7%, p=0.016). Conclusions: CBR3 genetic variants may influence the pharmacokinetics of doxorubicin and its major metabolite doxorubicinol. Inter-ethnic differences in frequencies of CBR3 genetic variants exist and may account for differences in pharmacokinetics and pharmacodynamics of doxorubicin between different populations.

No significant financial relationships to disclose.
Final results of a phase I clinical trial of the bioreductive drug RH1
Authors: Danson S. , Johnson P. , Ward T. , Dawson M. , Denneny O. , Watson A. , Jowle D. , Sharpe P. , Dive C. , Ranson M.
Published: March 2020

2514

Background: RH1 (2,5-diaziridinyl-3-[hydroxymethyl]-6-methyl-1,4-benoquinone) is a novel bioreductive drug which is activated by the two electron reductase, DT-diaphorase (DTD) (NAD(P)H:quinone oxidoreductase). DTD is over-expressed in many tumors relative to normal tissue, especially lung, breast and colorectal tumors, so it is hoped that RH1 is selectively activated in such tumours with minimal normal tissue toxicity. Methods: Under the auspices of the CR-UK Drug Development Office, a dose escalation phase I trial of RH1 was performed at two centres. The primary objective was to establish the maximum tolerated dose (MTD) of RH1 given as a 10–30 minute infusion for 5 days every 3 weeks. Toxicity was assessed according to NCI CTC V2.0 criteria. Secondary objectives were to determine RH1 pharmacokinetics (PK), DNA cross-linking in peripheral blood lymphocytes and tumour cells, DTD activity in tumour, NQO1 polymorphism status and document any anti-tumour activity. Results: Eighteen patients of WHO performance status 0–1 with advanced refractory solid malignancies were enrolled. 14 patients were male and eight patients had colorectal adenocarcinoma. The MTD was 1,430 mcg/m2/day and the dose-limiting toxicity (DLT) was bone marrow suppression. DLT occurred in both patients treated at 1,905 mcg/m2/day: one developed grade 3 thrombocytopenia with haemorrhage and grade 3 anaemia; the other died from neutropenic sepsis. Other clinically significant drug-related toxicities included moderate phlebitis and fatigue. Plasma PK analysis on days 1 and 5 in cycle 1 showed rapid plasma clearance of RH1 (t1/2=12.3 mins) with AUC increasing proportionately with dose. The comet-X assay showed increases in DNA inter-stand cross-linking in peripheral blood lymphocytes treated at 200 mcg/m2/day or above. These cross-links were detectable within one hour of commencement of dosing and reached a maximum of 30%. DNA cross-linking after RH1 was seen in tumour samples, even with low levels of DTD. One patient was homozygous for the NQO1 polymorphism. No objective tumour responses were recorded. Conclusions: The MTD of 1,430 mcg/m2/day is the dose recommended for phase II clinical trials. Tested doses were associated with DNA cross-linking in both peripheral blood lymphocytes and tumour cells.

No significant financial relationships to disclose.
A phase I study of MGCD0103 given as a twice weekly oral dose in patients with advanced leukemias or myelodysplastic syndromes (MDS)
Authors: Lancet J. E. , Nichols G. , Assouline S. , Ward M. , Burton M. , Mintz M. , Rousseau C. , Kalita A. , Brabant P. J. , Odenike O.
Published: March 2020

2516

Background: Small molecule inhibitors of histone deacetylase (HDAC) have emerged as novel anticancer agents. MGCD0103 is an oral isotype selective small molecule inhibitor of HDAC. Methods: Eligible patients had relapsed/refractory (RR) AML or MDS (or newly diagnosed disease in patients >60 yrs, ineligible for induction chemotherapy); RR ALL; RR CML. MGCD0103 was administered 2x/weekly for 3 weeks, with no recovery period between cycles. Patients with stable disease or better were eligible to continue therapy indefinitely. Results: Patients have been treated at 4 dose levels: 40, 53, 66 and 83 mg/m2/day, including 19 enrolled patients with the following demographics: M:F = 15:4, median age (range) = 75 (52–83), ECOG 0:1:2 = 5:12:2, diagnosis of RR MDS= 7, RR AML or RR ALL=8: untreated AML or MDS = 4. Cytogenetics (n=12): Diploid = 3, Complex = 4, Del 7 = 3, Del 12 = 1, Del Y = 1. Nineteen patients are evaluable for safety. A total of 38 cycles have been administered with a median of 2 per patient (range, 1–6); 13 patients have completed =2 cycles. One patient experienced grade 3 fatigue at 53 mg/m2 and 2 patients had grade 3 weakness/fatigue at 83 mg/m2 (exceeded the Maximum Tolerated Dose [MTD]) Non-dose-limiting toxicities included lower grade fatigue, diarrhea, and nausea. Plasma PK in 14 patients revealed t 1/2 of 7–12 hr, tmax 0.6- 1 hr, and a dose proportional average Cmax of 155 ng/mL at 40 mg/m2 and 225 ng/mL at 53 mg/m2. Significant inhibition of whole cell total HDAC activity within PBMC was observed in a majority of patients, at all dosing levels. Four patients have experienced stable disease. Conclusions: MGCD0103 has been well-tolerated in patients with advanced leukemias or MDS. MTD has been reached, and the recommended phase II 2x/week dose is being confirmed. At all dose levels, significant HDAC inhibition was observed.

[Table: see text]
Phase I study of PM00104, a novel cytotoxic anti-cancer drug, administered to patients with advanced solid tumors or lymphoma
Authors: Gallerani E. , Yap T. A. , Lopez A. , Coronado C. , Shaw H. , Florez A. , de las Heras B. , Cortes-Funes H. , de Bono J. , Paz-Ares L.
Published: March 2020

2517

Background: PM00104 (Zalypsis [Z]) is a novel synthetic alkaloid related to the marine compounds Jorumycin and the family of Renieramycins. Preliminary analyses of its mechanisms of action suggest effects on the cell cycle, DNA binding properties, as well as transcriptional inhibition. Z has shown antitumor activity in vitro (IC50 =10-8 M) and in xenografts models, and an acceptable toxicology profile. Methods: Patients (pts) with advanced cancers or lymphoma were enrolled to determine the safety, tolerability, maximum tolerated dose (MTD), recommended dose (RD), pharmacokinetics (PK) and antitumor activity of Z administered as a 1- hour IV infusion every 3 weeks. Sequential cohorts of 3–6 pts received the following doses: 0.225, 0.45, 0.9, 1.8, 3.0 and 3.6 mg/m2. Results: 27 pts were treated (19 M; median age: 58, range: 39–79; ECOG PS =2). 2 of 6 pts at 3.6 mg/m2 experienced dose-limiting toxicities (DLT), defining the MTD; 1 pt with colorectal cancer with extensive liver metastases developed grade 4 thrombocytopenia and neutropenia, grade 3 transaminase elevation, and grade 4 troponin I rise without cardiac symptoms or electrocardiogram alterations. These toxicities all resolved. Another pt with adrenal carcinoma developed grade 3 nausea, vomiting and asthenia despite anti-emetics, which resolved with dexamethasone 2 days after infusion. Other toxicities were =grade 2 and included: nausea, vomiting, transient transaminase elevation and asthenia at doses =0.225 mg/m2 and myelosuppression (neutropenia and leukopenia) at doses =3.0 mg/m2. No DLTs were seen in 6 pts at a dose of 3.0 mg/m2, establishing this dose as the RD. 3 more pts were added to complete the expanded cohort at this RD. To date, 2 pts have SD lasting >3 months. PK data indicate that drug AUC and Cmax increase proportionally across the dose range until doses =3.0 mg/m2, when a disproportionate increase in AUC and Cmax was seen. Z has a long half-life (=50 hours) and wide volume of distribution (Vss ∼500 l/m2). Conclusions: Z has a favorable safety profile. The MTD on this schedule has been established at 3.6 mg/m2. Alternative schedules are being evaluated to try and increase dose intensity.

No significant financial relationships to disclose.
Calculating carboplatin doses using the 4-variable modification of diet in renal disease (4-v MDRD) estimate of glomerular filtration rate (GFR) in the Calvert formula
Authors: Poole S. G. , Dooley M. J. , Rischin D.
Published: March 2020

2521

Background: The dose of carboplatin is usually calculated using the Calvert formula, with various bedside estimates utilized rather than directly measured GFR. The 4-v MDRD equation (Levey AS, et al. Ann Intern Med 2006; 145: 247–254) is now advocated as the routine method of estimating renal function from serum creatinine in all patients. Adoption in clinical practice is occurring despite lack of validation in oncology patients. The aim of this study was to compare carboplatin doses derived from the Calvert formula using measured GFR and the estimates of the 4-v MDRD equation and other established estimates. Methods: GFR was determined using technetium-99m diethyl triamine penta-acetic acid (Tc99mDTPA) clearance. Serum creatinine (Jaffe method) was measured and GFR estimates calculated using 4-v MDRD, Cockcroft and Gault formula (CGF), Wright, Martin, and Jelliffe (JF) formulae. Carboplatin doses were calculated using the Calvert formula, targeting an area under the curve of 7mg.ml- 1.min-1. Results: GFR was measured in 510 adult oncology patients (323 male, 187 female, mean age 63 years, range 17–87 years, mean GFR 84 mL/min, range 16–205 mL/min). The mean (range) carboplatin dose was 765 mg (287–1,610 mg), 681 mg (237–1,306 mg), 674 mg (249–2,044 mg), 721 mg (261–1,536mg), 741 mg (261–2,128mg), 620 mg (244- 1,329 mg) for measured GFR, 4-v MDRD, CGF, Wright, Martin, and JF formulas respectively. The accuracy (% within 20% of ‘true’ dose) was 58%, 63%, 73%, 72% and 49% for 4-v MDRD, CGF, Wright, Martin, and JF formulas respectively. Carboplatin doses derived using the 4-v MDRD estimate of GFR become increasingly less accurate with increasing GFR (see table ). All other formulas performed similarly. Conclusions: The 4-v MDRD equation resulted in an imprecise estimation of carboplatin doses with the degree of variability dependant on the level of renal function.

[Table: see text]

No significant financial relationships to disclose.
Identification of predictive markers to differentiate ixabepilone from paclitaxel activity in ER-negative breast cancer patients
Authors: Wu S. , Chasalow S. , Lee H. , Xu L. , Paul B. , Mokliatchouk O. , Symmans W. F. , Zerba K. E. , Pusztai L. , Clark E.
Published: March 2020

2525

Background: Ixabepilone (BMS-247550) is a microtubule stabilizing agent with demonstrable therapeutic value in taxane- refractory breast cancer (BC) patients. Biomarkers to predict either ixabepilone or paclitaxel activity in BC patients have previously been reported. However, markers that differentiate response to the two agents have yet to be identified. This study sought to discover predictive markers that will enable patient selection to differentially enhance response to ixabepilone or paclitaxel in ER-negative (ER-) patients. Materials and Methodologies: Pre-treatment gene expression profiles were generated for 62 ER- patients treated with ixabepilone in clinical study CA163080, and 51 ER- patients treated with T/FAC (paclitaxel and fluorouracil-doxorubicin-cyclophosphamide) in clinical study MDA133. Biomarkers differentially predictive of complete pathological response in breast were identified through gene set enrichment analysis (GSEA) or classification by threshold gradient descent (TGD). Gene knockdown by siRNA was used to study some of these candidate markers. Results: Four candidate models that differentiate response to ixabepilone treatment and taxane-containing therapy were identified. Two of the models, found by GSEA, are based on expression levels for single microtubule-related genes: transforming, acidic coiled-coil containing protein 3 (TACC3) and chromosome condensation protein G (HCAP-G). The potential of HCAP-G as a differential marker was supported by siRNA studies. Two of the models, found by TGD, are based on expression levels for 26 and 20 genes. Areas under the ROC curves for the models applied to each study separately are given in the table . Conclusions: We have identified four predictive models that differentiate response in a clinical trial of ixabepilone from that in a trial of T/FAC. A clinical trial is under way to further evaluate their utility for differentiating response to ixabepilone- and taxane-containing regimens.

[Table: see text]

[Table: see text]
Relationship between glutathione-S-transferase P1 Ile105Val polymorphism and docetaxel neurosensory toxicity
Authors: Mir O. , Alexandre J. , Tran A. , Rabillon F. , Girre V. , Dieras V. , Pons G. , Treluyer J. M. , Goldwasser F.
Published: March 2020

2527

Background: Glutathione-S-transferases (GST) regulate the cellular response to oxidative stress and various anticancer agents. We recently underlined the importance of oxidative stress in the side effects of taxanes [Alexandre et al,J Natl Cancer Inst 2006;98:236–44], and investigated the relationship between the GST isoforms M1, T1 and P1 genes polymorphisms and docetaxel-induced peripheral neuropathy. Methods: We retrospectively analysed GST polymorphisms in a cohort of cancer patients (pts) treated with docetaxel and entered in a clinical trial database. A clinical neurologic evaluation (according to the NCI-CTC v2.0) was performed at baseline and at each treatment cycle. The GST M1 (null genotype), GST T1 (null genotype), and GST P1 (Ile105Val and Ala114Val) polymorphisms were determined using PCR, followed by either sequencing or RFLP techniques. The relationship between GST polymorphisms and grade = 2 neurosenrory toxicity (NST) as primary endpoint was studied, using the Chi2-square (with Yates correction) and the Fischer’s two-tailed exact test. Results: Fifty-eight pts were included : F/M: 29/29; median age: 61 years (range: 47–75). Primary tumor: 27.6% breast, 27.6% prostate, 24.1% lung and 20.7% other cancers. Pts received docetaxel 75–100 mg/m2 given as single-agent. A total of 261 cycles were administered (median/pt: 4, range 2–12). Ten pts developed grade = 2 NST. Twenty-seven pts (47%) were homozygous for the GST P1 105Ile allele, and 31 pts were either homozygous or heterozygous for the GST P1 105Val allele. Grade = 2 NST was significantly more common in pts with GST P1 105Ile/105Ile genotype (8/27 pts, 30%) compared with patients with 105Ile/105Val or 105Val/105Val GSTP1 genotype (2/31 pts, 6,5%; P = 0.047). Pts who were genotyped as GST P1 105Ile/105Ile had a higher risk of developing a grade = 2 NST than did those with other GSTP1 genotypes (OR 6.11; 95% CI, 1.17–31.94; P < 0.05). Conclusions: We found a significant correlation between GST P1 105Ile homozygous genotype and the development of a docetaxel-induced peripheral neuropathy.

No significant financial relationships to disclose.
Evaluation of pemetrexed (PEM) dosing paradigms using exposure-response relationships (ERRs)
Authors: Latz J. , Claret L. , Symanowski J. , Falcoz C. , Bruno R. , Smith I. , Calvert H.
Published: March 2020

2530

Background: PEM is primarily renally eliminated and dosed based on body surface area (BSA). A direct relationship exists between creatinine clearance (CrCL) and PEM clearance (CL [and therefore exposure, AUC]). BSA-dosing has been increasingly challenged, especially for renally cleared drugs. ERRs and dosing paradigms (BSA, fixed, renal-based) were evaluated in silico for PEM. A superior dosing strategy is dependent on the strength of the ERR and results in decreased between-patient variability in both AUC and clinical outcome. Methods: An established pharmacokinetic model was used to estimate AUCs from CrCL for patients in malignant pleural mesothelioma (MPM) and NSCLC studies where patients with CrCL =45 mL/min received PEM 500 mg/m2 q3w. AUC was evaluated as a predictor of clinical outcome (survival, TTPD, toxicities) to identify ERRs. Efficacy models included previously identified prognostic factors and inverse of mean AUC over the treatment period as covariates. Probability of toxicity was modeled as a function of AUC at each cycle. Simulations were performed for statistically and clinically significant ERRs to compare distributions of outcome for the 3 dosing paradigms. Results: AUC was individually significant (or borderline) in the survival and TTPD ERRs for MPM (PEM + cisplatin; N=226) and NSCLC (PEM; N=258). AUC was independently significant only for the NSCLC TTPD ERR. The distribution of simulated TTPDs for the 3 dosing paradigms were similar (see table ). Model-predicted increase in incidence of Grade 3–4 hematologic toxicities with increasing AUC was statistically but not clinically significant. Graphical review of nonhematologic toxicities showed trends by AUC. The incidence of death due to drug-related toxicities was too sparse for modeling. Conclusion: Neither renal-based nor fixed dosing provides a clinical benefit over BSA-based dosing for patients with CrCL =45 mL/min based on ERRs and simulations using AUCs estimated from CrCL.

[Table: see text]

No significant financial relationships to disclose.
Patterns of HER-family receptor dimerization in trastuzumab susceptible and trastuzumab resistant cell lines
Authors: Dua R. , Nhonthachit P. , Rinehart C. , Arteaga C. L. , Nahta R. , Esteva F. J. , Winslow J. , Bates M. , Petropoulos C.
Published: March 2020

2533

Background: HER2 overexpression is associated with accelerated disease progression and poor prognosis in breast cancer. Trastuzumab, a monoclonal antibody targeting the extracellular domain of HER2, is effective in the treatment of metastatic breast cancer. However, most patients treated with trastuzumab eventually develop clinical resistance. To investigate the role of HER-family receptors in trastuzumab resistance, we measured HER-family receptor expression, dimerization, and phosphorylation in trastuzumab susceptible and resistant cell lines. Methods: Cell lysates from trastuzumab susceptible and resistant BT474 and SKBR3 cell lines were obtained from the Arteaga and Esteva laboratories. Proximity-based, multiplexed assays were used to detect and quantify HER1, HER2, and HER3 expression and phosphorylation levels, as well as HER1/HER1, HER1/HER2, HER1/HER3, HER2/HER2, and HER2/HER3 dimers. Samples were incubated with a mixture of HER specific antibodies conjugated either with fluorescent reporter tags (eTags), or biotin, which binds a reporter tag releasing agent (chemical scissor). Reporter molecules are released based on proximity to the scissor in a photochemical reaction and separated by capillary gel electrophoresis. Results: In comparison to trastuzumab susceptible parental cell lines, both SKBR3 and BT474 trastuzumab-resistant cell lines displayed upregulated HER1 expression. Resistant BT474 cell lines exhibited markedly increased levels of HER1/HER2 heterodimers. Increases in HER2 phosphorylation in the trastuzumab resistant SKBR3 cell line were observed, consistent with previous studies implicating trastuzumab in the induction of HER2 phosphorylation. Total HER2 and HER3 levels were similar in trastuzumab susceptible and resistant BT474 cell lines. Conclusions: The development of trastuzumab resistance in these cell line models correlated with HER1 expression and the appearance of HER1:HER2 dimers. Since signaling initiated by such heterodimers is ineffectively antagonized by trastuzumab, these data suggest that selection for proliferative signaling mediated by HER1:HER2 dimers may represent a mechanism of trastuzumab resistance in breast cancer.

No significant financial relationships to disclose.
A genomic approach to identify mechanisms associated with chemotherapy resistance
Authors: Riedel R. F. , Porrello A. , Chenette E. , Potti A. , Nevins J. R. , Febbo P. G.
Published: March 2020

2534

Background: Gene expression profiling has shown an ability to predict chemotherapeutic response (Potti et al. Nature Medicine 2006). Building on this work, we used a genomic strategy to explore the biology associated with the development of chemotherapy resistance and sought to determine if disease context impacted results. Methods: Gene set enrichment analysis (GSEA) was performed on expression data for NCI60 cell lines sensitive and resistant to specific chemotherapeutic agents (adriamycin, cyclophosphamide, docetaxel, etoposide, 5-fluoruracil, paclitaxel, topotecan). GSEA was additionally performed on a series of lung cancer cell lines with a defined sensitivity to cisplatin and docetaxel. Sensitive and resistant cell lines with individual mean and confidence intervals greater than 1SD from the mean across all samples were included. Adjusting for multiple hypothesis testing, gene sets with a false discovery rate (FDR) <0.25 were deemed statistically significant. In the discovery mode, gene sets with a nominal p-value <0.05 were also of interest. Finally, overlapping gene sets between agents were assessed. Results: Statistically-significant gene sets, representing biologic pathways associated with resistance, were identified for the various chemotherapeutic agents (i.e., cell death, erbb3, and bad pathways associated with docetaxel resistance). No gene sets with FDR <0.25 or nominal p-value <0.05 were common to all drugs. In assessing disease specific resistance, 22 lung cancer cell lines for cisplatin (15 sensitive, 7 resistant) and 14 lung cancer cell lines for docetaxel (10 sensitive, 4 resistant) were analyzed. GSEA identified the bcl-2 pathway (p<0.002) to be associated with cisplatin resistance. In contrast, the proteosome (p=0.01) and akt (p=0.02) pathways were associated with docetaxel resistance. Pathways involved in the production of V-H+-ATPase were enriched in cisplatin and docetaxel resistant lung cancer cell lines suggesting a global mechanism of resistance. Conclusions: These results support the use of a genomic approach to identify unique drug-specific and global therapeutic targets associated with the development of chemotherapy resistance. Interestingly, disease context appears important in identifying novel targets.

No significant financial relationships to disclose.