bluuebunny/crossref_metadata_2025 huggingface dataset:
Extranodal NHL- A retrospective review of clinico-pathologic features and outcome and comparison with nodal NHL. Single institution experience years: 1988–2004
Authors: Lal A. , Adil S. , Masood N.
Published: March 2020

17542

Background: Non-Hodgkin’s lymphoma (NHL) arising in an extra nodal (EN) site is not uncommon and its natural history and treatment is clearly characterized in the literature. Data on EN-NHL and comparison with N-NHL with relation to survival and prognostic factors is scarce in our part of the world. The primary objective of this study was to analyze the anatomic distribution, clinical features and outcome of DLBCL patients according to the primary site with applicability of International Prognostic Index (IPI). Methods: From 1988 to 2004, 557 patients were analyzed for the clinico-pathologic characteristics, treatment outcome and prognostic factors affecting overall survival. Results: Median age was 48.7 ± 15.3 years ; the M: F ratio was 2:1. The distribution according to the primary site was: lymph node, 322 cases (58%) of these 145 cases (44%) stage IV, 76 cases (23%) Stage III, 60 cases (18%) stage II and 47 cases(15%) stage I ; and EN sites, 235 (42%), including GIT (44%) followed by upper aerodigestive tract (19%), bones (08%), spine (05%), and 3% each as breast, CNS, testis,lungs. The median survival rate was 4.8 and 6.3 years in NL and ENL respectively vary according to primary site/stage of the lymphoma. In the univariate analysis age less than 60 years, early stage I-II, extra nodal involvement primarily gastric or bone, 0–1 extra nodal site, 0–1 PS, lack of B symptoms, normal LDH level has been associated with good prognosis. In the multivariate analysis age, PS, stage and level of LDH were the main variables to predict OS; no nodal or extranodal site maintained their prognostic value. Conclusion: Our data correspond with series from west increasing incidence extranodal lymphoma due to improved diagnostic techniques and superior results with chemotherapy by preserving the organ. Few patients with bowel obstruction or cord compression lymphoma required surgery for diagnosis or relief of symptoms. There is significant difference from western data in histologies DLBC-NHL is the most common histologies in our study. Overall survival patients with EN-NHL were similar to nodal NH-Lymphoma but largely depended on IPI.

No significant financial relationships to disclose.
The use and correlation with flow cytometry of fluorescent molecular beacons in real time PCR of IgH gene rearrangements for evaluation of minimal residual disease in multiple myeloma
Authors: Kara O. , Yigin A. , Sahin B. , Paydas S.
Published: March 2020

17552

Background: At present, the prognostic value of the amount of residual tumor cells in PB, BM or stem cell harvests and its changes over time is stil not clear. Also the advent of new therapeutic approaches to multiple myeloma made necessary the introduction of novel methods for detection of minimal residual disease. Among others approaches residual disease can be detected by using flow cytometry. The aim of the present study was to evaluate a real time PCR test for the IgH gene using alellespecific molecular beacons as fluorescence probes to quantify residual disease and also correlate flow cytometric detection of plasma cells in MM patients during followup after treatment with high dose of chemotherapy or standart chemotherapy. Methods: After clinical diagnosis of 17 MM patients, the CDR1, CDR2 and CDR3 regions of the IgH gene were analysed and sequenced to identify its clonal nature. Unique sequences of the clonal IgH rearrangement were used to design specific molecular beacon probes for each MM patient. We have also examined the co-expression of CD19, CD38, CD45, CD56, and CD138 molecules in cells of bone marrow aspirates in patients with multiple myeloma by flow cytometry. Results: The active disease had been accepted of whom plasma cell infiltration ratio was over 10% in bone marrow and also of whom labeled by CD38 and CD138 by FCM. The detection of the MRD was positive in 13 patients by RT-PCR, respectively. The infiltration ratio was correlated with CD138 expression (p = 0.009) and RT-PCR detection of plasma cells (p = 0.006) and also significant correlation had been found between RT-PCR detection and CD138 expression respectively. No any correlaton was found between other surface antigens (CD38, CD45, CD56). Conclusion: Our results indicated that real time PCR with specific molecular beacons provides a feasible, accurate and reproducible method for the determination of minimal residual disease in MM. By FCM only CD138 expression may have been used as disease marker in addition of the RT-PCR detection.

No significant financial relationships to disclose.
Hodgkin’s lymphoma in the elderly veterans: We need to do better
Authors: Al Sayyed M. H. , Maken S. F. , Safa M. M. , Nahleh Z. A. , Pancoast J. R. , Muhleman A. F. , Jazieh A. R. , Komrokji R. S.
Published: March 2020

17554

Background: Recent studies demonstrated worse outcome in elderly patients diagnosed with Hodgkin’s Lymphoma (HL). The purpose of this study was to compare the presenting features, treatment, and outcome of elderly patients with HL within the Veteran Affairs Health care system (VA). Methods: This was a retrospective analysis. The VA Central Cancer Registry (VACCR) database was used to identify patients with HL diagnosed between 1995 and 2005. There are approximately 120 VA medical centers diagnosing and/or treating patients with cancer. Data are entered by tumor registrars at the VA medical centers. This site aggregates the data collected by the medical centers’ cancer registries. Data was extrapolated and analyzed using bio-statistical software SPSS. Results: We analyzed 1009 patients in the Veteran Affairs (VA) database with the diagnosis of Hodgkin’s lymphoma. Patients were divided into two groups according to age, elderly group (> 60 years) (n = 383) and young group (Age ≤ 60 years) (n = 626). The median age of presentation in elderly patients was 70 years; the most common stage at diagnosis was Stage IV. The most common histological subtype was nodular sclerosis and bulky disease was present in 89 (23.2%) of patients. Out of 383 patients 237 (62%) were treated with chemotherapy, 88 patients (23%) received radiation. Compared to patients ≤ 60 years of age, elderly patients were less likely to be current smokers, more likely to be caucasian, all other baseline characteristics were not statistically significantly different. Less elderly patients received chemotherapy compared to young patients (62% versus 74%) (P-value 0.004). No difference was observed in radiation therapy. The 5 year Overall survival in the elderly was 27 % compared to 70% in patients aged 60 or less. (P-value <0.005). Conclusions: Outcome of HL among elderly veterans is poor. Elderly patients received less chemotherapy treatment. Novel strategies to better deliver treatment in elderly patients with HL are warranted.

No significant financial relationships to disclose.
Primary mediastinal large B-cell lymphoma: Treatment outcomes in 43 patients
Authors: Mello C. L. , Feher O. , Lima V. C. , Valadares C. , Soares F. A. , Carvalho A. L.
Published: March 2020

17556

Objectives: Primary mediastinal B-cell Lymphoma (PMBL) is recognized as a separate entity in the WHO classification. Treatment for PMBL is based on a combination of conventional dose chemotherapy, high dose chemotherapy and radiation therapy. The best strategy is still undefined. We conducted a retrospective analysis of patients with PBML to identify clinical prognostic factors. Methods: A retrospective analysis of 43 patients treated at Hospital do Cancer AC Camargo, Sao Paulo, Brazil, between 1989 and 2004. All patients had previous diagnosis of diffuse large B-cell lymphoma, with positive CD20 on neoplastic cells. A predominant anterior mediastinal lesion should be present. Induction chemotherapy regimens were grouped in first generation (CHOP/CHOP-like), third generation (PromaceCytabon/MACOP) and other (pediatric regimens, COP). Results: Age ranged from 16 to 82 years-old, 30 females and 13 male. Age < 35 yo was associated with a better prognosis (5 years OS - 56% × 34%, p = 0.048). Among clinical variables, female gender, stage IA-IIB, IPI 0–1, normal LDH, absence of mediastinal bulky disease were associated with better prognosis, although not statistically significant. Response rate to first generation regimens was: 37% CR (11/29), 24% PR (7/29) and 24% PD (7/29). Four patients were treated with Third generation regimens with 2 CR and 2 PR. 20 out of 25 patients with PR or CR to first line chemotherapy received mediastinal radiation therapy. More than 65% of patients had a follow up of 5 years or more. With a median follow up of 22.3 months, projected 5 year OS was 47% and for the responders the median PFS was 8,4 months. No difference in OS and PFS was observed among the three chemotherapy groups. Conclusion: Our analysis showed that response rate to first line regimens was around 60% and 25% of patients were primarily refractory to CHOP regimen. Age younger than 35 years old was associated with a better prognosis. 5 years overall survival was 45% and is in accordance with the literature. Although recent studies have demonstrated biological similarities between PMBL and Hodgkin’s Lymphoma, the prognosis of PMBL is less favorable than HL. Better understanding of the disease will help in developing more appropriate therapeutic strategies for PMBL.

No significant financial relationships to disclose.
Needs satisfaction for cancer patients in an ambulatory chemotherapy service (ACS). An exploratory study
Authors: Wbanet S. , Cayuela A. , Bernal L. , Valderrama B. , Valero M. , Ruiz M. , Iglesias M. , Iglesias L. , Moreno J. , Casas A. M.
Published: March 2020

18607

Background: ACS are currently the main way for the delivery of Chemotherapy to cancer patients. Aproximately half of them require intravenous chemotherapy and many need two or three separate lines of treatment during their clinical career. How these treatments are delivered will affect their health and quality of life. Appraisal of needs along chemotherapy process enable to identify problems related to health satisfaction status and quality of health care provided. Methods: We have evaluated the perceived needs of patients receiving ambulatory chemotherapy through an “ad hoc” questionnaire of 49 items exploring the following areas: treatment related symptoms, information and communication/interpersonal interaction, psychological, social and spiritual support, economical needs and management of health service. Results: 141 consecutive patients attending the ACS one year after its set up were evaluated. Demographics (age, gender and income), diagnosis and types of Chemotherapy (Adjuvant vs. Palliative) were considered for interrelationships. The most frequent tumors were breast and colorectal carcinoma. More than 50% of the patients reported having controlled symptoms (pain, nausea and vomiting, mucositis etc.). Information needs related to their illness and treatment were satisfied to more than half of the sample, but reported a need for more information about diagnostic procedures and prognosis; 48% perceived shortcomings in communication with their clinicians and 50% believed they received little advice about how cancer affects their sexual behaviour. Most patients (53%) reported not to need pshycological support neither for their family (60%) nor themselves, 83% not needing spiritual support and 50% were satisfied with the economical support they received. With respect to the facilities’ structure, 50% perceived enough intimacy in doctor’s offices and Chemotherapy Room and 64% felt comfortable in the waiting room but 72% perceived waiting too long before being attended by doctor’s and nurses. Conclusions: younger patients and breast cancer diagnosis are among the people that express more unsatisfaction related to the accomplishment of their needs (p < 0.05).

No significant financial relationships to disclose.
Is bisphosphonate-induced osteonecrosis of the jaw (BONJ) an ischemic manifestation of pseudo-osteopetrosis?: Biochemical, densitometric, and imaging evidence
Authors: Kelleher F. C. , McKenna M. , Collins C. , Brady G. M. , Crown J. P.
Published: March 2020

18621

Background: BONJ is an increasingly recognised but poorly understood complication of long-term B therapy. We hypothesise that the pathophysiology is ischaemic, and is due to an osteopetrosis (OP)-like lesion which causes progressive constriction of maxillo- mandibular perforating arterial foramina. Support for this comes from the observation that the osteoclast ruffled border is functionally impaired in osteopetrosis and structurally lost with B therapy. Cases of maxillomandibular osteomyelitis and avascular necrosis complicating OP exist. Additionally the blood supply of the jaw relies partly on small periosteal perforators. Methods: Bone mineral density (BMD) of 2 female patients (pt) with BONJ was investigated with dual energy x-ray absorbptiometry. Serum creatinine BB-isoenzyme (CK-BB) was determined. Serum acid phosphatase was assessed for patient 1. Bone biochemistry; (serum calcium, phosphate, parathyroid hormone, bone alkaline phosphatase, 25(OH) vitamin D, osteocalcin (intact), N. Ter.procoll1, CTX-1) was assessed. Multislice CT with orthoradial reconstruction was performed on pt 1. Results: CK-BB isoenzyme was increased at 21% and 20% in patients 1 and 2 (upper limit normal 2%). DEXA scan patient 1 results: BMD, lumbar spine; T score 4.9 (152%), Z score 5.4 (160%), femur; T score 2.2 (128%) Z score 2.5 (134%). DEXA scan patient 2 results: BMD lumbar spine; T score 0.2, Z score 1.2, femur; T score 1.2, Z score 1.8. Serum acid phosphatase in pt 1 was normal. Orthoradial reconstruction of the mandible in pt 1 showed absence of lateral lingual canals in affected hemi mandible.Mental foramen and inferior alveolar canal were not constricted. Conclusion: The increased BMD and CK BB isoenzyme seen in our pts are characteristic features of osteopetrosis. The normal acid phosphatase is explained by bisphosphonate-induced osteoclast apoptosis a phenomenon that does not occur in osteopetrosis. Orthoradial reconstruction of the mandible provided radiological correlation. The data was found to be consistent with the hypothesized disease process.

No significant financial relationships to disclose.
Gastrointestinal stromal tumors (GISTs): Our experience in the management of 26 patients
Authors: Maiello E. , Tozzi L. , Latiano T. P. , De Bonis A. , Bisceglia M. , D’Addetta C. , Di Sebastiano P. , Nanni L. , Valori V. M. , Piano A.
Published: March 2020

19506

Background: GISTs represents 0.1–3% of gastrointestinal neoplasms; most cases occur in people 40 to 80 years old, and are more common in men than in women. More than half of all GISTs patients present with locally advanced, recurrent, or metastatic disease (mainly to liver or peritoneum). Methods: From January 2001 to December 2005 we observed in our Institution 26 pts with GISTs; all were CD117+.The main characteristics of these pts were as follows: median age: 61 yr; sex (F/M): 14/12; primary tumor site: stomach 13 (50 %), small intestine 7 (27 %), rectum 1 (4 %), retroperitoneum 4 (15 %), liver 1 (4 %); first symptoms: epigastric pain 10 (38 %); abdominal pain 4 (15 %); hematemesis 1 (3 %); anemia 5 (18,5 %), palpable abdominal mass 1 (3 %), defecation disorders 2 (7 %); melena 3 (11 %); rectal bleeding 1 (3 %); ascites 1 (3 %). Results: Radical surgical resection was performed in 21 patients (81 %), 2 patients received a debulking surgery, and no postoperative mortality or major complications were observed; 3 pts showed an advanced disease. Seven pts (33%) developed recurrence, local or at distance, and the median time to relapse was 7,5 months (range 2–11). One pt with advanced disease dead before any treatment and 11 pts received Imatinib (I), at the standard dose (400 mg/d), starting from the date of diagnosis of advanced disease or metastatic relapse and given until development of intolerance or progressive disease. Main toxicities of I included: neutropenia G3 (4%), skin rash (4%), periorbital oedema (4%). We achieved 3 PR (lasting 12+, 21+ and 40 mos), 6 SD and 2 PD. With a median follow-up of 30 months, all but three pts (dead for progressive diseases) are alive. The overall survival rate was 88.5 % and the median overall survival was 16 mos (range 1–49). Conclusions: Surgical resection remains the only effective treatment for GISTs. However, in pts with advanced or relapsed disease treatment with I is effective with an high disease control rate.

No significant financial relationships to disclose.
Prediction of response to neoadjuvant chemotherapy in breast cancer
Authors: Hannemann J. , Halfwerk H. , Velds A. , Loo C. , Rutgers E. , Rodenhuis S. , Van de Vijver M.
Published: March 2020

20005

Background: Neoadjuvant chemotherapy is increasingly employed in operable breast cancer. Our initial studies on a cDNA array platform failed to identify gene expression patterns predicting response to neoadjuvant chemotherapy in breast cancer patients (J Clin Oncol 23:3331, 2005). Now we included more patients and used oligo microarrays. Methods: Patients with operable or locally advanced breast cancer were included in a randomized phase II study or received neoadjuvant chemotherapy off protocol. All except 7 patients began chemotherapy with 3 courses of dose-dense adriamycin and cyclophosphamide (ddAC) and response was evaluated by MRI. Patients with a response and a HER2-positive tumor were then randomized between either 3 additional courses of ddAC or six weekly courses of carboplatin, paclitaxel and trastuzumab (CPT). Patients without response were switched to CPT. Patients with HER2-negative tumors were randomized between 3 courses of either ddAC or capecitabine and docetaxel (CD). After evaluation, patients without response were switched to the alternative treatment arm. From all patients 14G core needle biopsies were taken before treatment and total RNA was isolated. Amplified mRNA was labeled and hybridized to 35k human oligo microarrays from our microarray facility. Results: So far, 77 patients have been included into the study. From 48 of these, good quality RNA from tissue with >50% tumor cells was isolated. 43 patients had received ddAC as initial chemotherapy; 32 of these had not been switched to another regimen. In a training set containing 11 pathological complete remissions (pCR) and 9 non-responders (NR) we could separate these groups by using 20 genes in a supervised classification and a 9-step cross validation. These results could be validated in an independent set of 11 samples (6 pCR, 5 NR). From 10 out of 11 samples, response status could be predicted correctly, independent from the treatment regimen. Although ER-positive tumors have a lower pCR rate than ER-negative ones, the steroid hormone receptors were not present in the classifier. Conclusions: We conclude that it should be possible to identify a reliable gene expression profile associated with response to adriamycin based neoadjuvant chemotherapy in breast cancer.

No significant financial relationships to disclose.
HER2 comparison with novel 3-gene marker sets for risk prediction of distant recurrence of breast carcinoma
Authors: Alsobrook J. , Hraber P. , Davis L. , Harris C. , Doherty P. , Hall B. , Williams T. , Hozier J.
Published: March 2020

20009

Background: The risk of distant recurrence in patients with invasive breast carcinoma (BrCa) is predicted imperfectly by currently used factors (e.g. nuclear grade, hormone receptor status), complicating adjuvant therapy decisions. In an effort to discover biomarkers for recurrence, we retrospectively analyzed BrCa recurrence and tumor gene amplification. Multi-gene marker sets were evaluated for predictive significance and accuracy. Because studies of HER2 as a suitable biomarker report positive and negative results, we compared our marker sets with HER2. Methods: 229 of 723 BrCa patients met inclusion criteria: invasive ductal carcinoma, no isolated local recurrence, minimum 4 years follow-up, and archived specimens adequate for fluorescent in situ hybridization (FISH) assay. Study endpoint was recurrence, or non-recurrence throughout follow-up. FISH was performed on formalin-fixed paraffin sections with probes for previously identified genomic regions. Signals were counted with a Metasystems workstation, and used in a “Prognostic Index” (PI) to categorize recurrence risk. One-sided Fisher’s exact test was used to assess increased incidence of distant recurrence in relation to PI and HER2, and increased incidence of HER2 amplification in relation to PI. Results: Two separate 3-gene marker sets were obtained that predict clinical outcome for hormone receptor + (HR+) and hormone receptor − (HR−) tumors. In HR+ samples a PI based upon regions surrounding CYP24, PDCP6IP, and BIRC5 was the best predictor of recurrence, while for HR- specimens the PI was based on the NR1D1, SMARCE1, and BIRC5 regions. The PIs are significantly associated with recurrence of HR+ and HR− tumors (OR = 9.52, 95% CI > 2.12, p = 0.0024; and OR = 12.3, 95% CI > 1.45, p = 0.0188, respectively). However, tumor HER2 amplification was not prognostic for recurrence (OR = 1.76, 95% CI > 0.77, p = 0.1395). Furthermore, there was no significant association of PI with HER2 status (OR = 1.56, CI > 0.57, p = 0.862). Conclusions: Prognostic Indices based on copy numbers of two 3-gene marker sets significantly predict clinical outcome of stage I-III invasive ductal carcinoma. HER2 amplification, included in our previous whole genome survey but not present in our final marker sets, had no prognostic value.

[Table: see text]
Characterization of a subset of CD4+ T cells expressing CD161 in cancer patients and healthy individuals
Authors: Iliopoulou E. G. , Karamouzis M. V. , Perez S. A. , Ardavanis A. , Baxevanis C. N. , Papamichail M. , Rigatos G.
Published: March 2020

20016

Background: CD161 is a glycoprotein expressed in >90% of NK and 25% of T cells in the peripheral blood of healthy individuals. Several NK receptors on T cells infiltrating tumors have been proven to negatively influence their effector function and therefore play a role in tumor escape. In this study, we investigated T cells expressing CD161 in the peripheral blood mononuclear cells (PBMC), tumor infiltrarting lymphocytes (TIL) or malignant effusions (ME) from patients with several types of cancer. Methods: Expression of CD161 in CD4+ or CD8+ (lacking CD56) T cells, was examined using four-colour flow cytometry. The proliferative capacity and potential cytokine production of purified CD4+CD161+CD56− cells, were studied after weak or strong stimulation, with or without costimulation, in the presence or absence of Interleukin-2 (IL-2). The possible regulatory function of activated CD4+CD161+CD56− cells on T cell allo-responses was also investigated. Results: CD4+CD161+CD56− T cells were significantly increased (P < 0.01) in TIL, either from tumor tissue (n = 8) or metastatic lymph nodes (n = 5), and ME (n = 25), compared to PBMC from both cancer patients (n = 36) and healthy individuals (n = 12). CD4+CD161+CD56− cells from all sources tested, have the same phenotypic characteristics: they comprise a memory T cell population (CD45RO+CD45RA−) expressing high CD28 and CD95 and low CD25, CD38 and HLA-DR. Co-stimulation via CD28 is important for induction of proliferation and production of large amounts of Th1 and Th2 cytokines (IFN-γ, TNF-a, GM-CSF, IL-4 and IL-10). Following co-stimulation, CD4+CD161+CD56− cells also exert a suppressive activity on autologous PBMC allo-responses. The latter effect does not require cell-to-cell contact and is mediated by soluble factors, including IL10, since neutralization of IL10 partially restored the immune response. Conclusions: CD4+CD161+CD56− cells represent a distinct memory T cell population that is significantly increased in TIL and ME in patients with cancer. These cells are capable of secreting large amounts of both Th1 and Th2 cytokines and might play an immunosuppressive role, mainly through IL-10 production, depending on the microenvironment in which they develop.

No significant financial relationships to disclose.
Elevated multidrug resistance-associated protein 4 (MRP4) expression in localized prostate cancer—A potential androgen regulated protein
Authors: Horvah L. G. , Ho L. , Kench J. G. , Allen J. A. , Scheffer G. L. , Stricker P. D. , Grygiel J. J. , Sutherland R. L. , Henshall S. M.
Published: March 2020

20022

Background: MRP4 is an ATP-binding cassette transporter and amphipathic anion efflux pump which transports prostaglandins, nucleoside analogues, glutamate and phosphate analogues. High MRP4 expression is prognostic of poor outcome in neuroblastoma and also correlates with MYCN amplification, suggesting regulation by this oncogene. Although MRP4 is known to be expressed in normal prostate epithelium, its expression in prostate cancer (PC) is undefined. This study aimed to define the pattern of expression of MRP4 in normal and malignant prostate tissue and assess the association with androgen exposure. Methods: 84 radical prostatectomy specimens from patients with clinically localized PC (22 neoadjuvant androgen ablation therapy, 62 no neoadjuvant treatment), 42 specimens of hyperplasia adjacent to PC and 16 cases of advanced PC were assessed for MRP4 expression using in situ hybridisation and immunohistochemistry. PC cell lines were assessed by immunoblotting. Results: There were significantly higher levels of MRP4 mRNA and protein expression in localized PC compared to hyperplasia (p=0.006). Conversely, MRP4 protein levels were significantly decreased in PCs treated with neoadjuvant androgen ablation therapy compared to cancers exposed to normal testosterone levels (p < 0.0001). There was also a trend towards decreased MRP4 expression in advanced PCs. Furthermore, immunoblotting revealed that MRP4 protein was more highly expressed in androgen-dependent (LNCaP) compared to androgen-independent (PC3/DU145) cell lines. In addition, in a panel of 14 normal human tissues only kidney and prostate tissue expressed MRP4 protein suggesting limited expression of MRP4 in human tissues. Discussion: Elevated MRP4 expression is found in malignant compared to benign prostate tissue while lower MRP4 expression is seen after androgen ablation suggesting that MRP4 may be an androgen-regulated gene. In addition, there is relatively little expression of MRP4 in normal tissues. These data suggest that MRP4 is important in the progression to PC and that it may be a potential therapeutic target.

No significant financial relationships to disclose.
Epidermal growth factor receptor expression correlates with histologic grade but not microsatellite instability in primary colon carcinomas
Authors: Rego R. L. , French A. J. , Foster N. R. , Sargent D. J. , Alberts S. R. , Windschitl H. , O’Connell M. J. , Thibodeau S. N. , Sinicrope F. A.
Published: March 2020

20023

Background: EGFR is a transmembrane receptor tyrosine kinase belonging to the ErbB family of proteins. Anti-EGFR antibodies have shown efficacy as monotherapy and can enhance cytotoxic chemotherapy in patients with advanced colorectal cancers. We studied EGFR expression in primary colon cancers and its association with, microsatellite instability (MSI), clinicopathological variables and patient survival rates. Methods: Archival colon carcinomas were studied from patients with high risk Dukes’ stage B2 (n = 78) and C (n = 230) patients enrolled in a prior adjuvant trial of 5-fluorouracil, leucovorin plus standard dose or high-dose levamisole (NCCTG 91–46–53). EGFR expression (anti-EGFR H11 antibody, DAKO) was analyzed by immunohistochemistry (IHC) and intensity (0–3+), extent and their product (weighted score) were determined. MSI was analyzed using BAT26 and expression of hMLH1, hMSH2 and hMSH6 proteins by IHC. Patients were censored at 5 years after randomization for DFS and censored at 8 years post study randomization for overall survival (OS) data. Results: EGFR protein expression was detected in 216 of 308 (70%) of Dukes’ B2 and C primary colon carcinomas from patients and localized to the plasma membrane. EGFR expression variables (intensity, extent, weighted score) were increased in poor/undifferentiated compared to moderate/well differentiated tumors (p ≤ 0.036). Similar EGFR expression levels were found in MSI-H (n = 27) versus MSS (n = 203) colon cancers and for other clinicopathological variables. EGFR intensity was more likely moderate (2+) or strong (3+) in Dukes’ C versus B2 tumors (p = 0.088). In addition, higher EGFR intensity (2+, 3+) showed a trend toward worse disease-free survival (5 yr DFS %: 62.2 vs. 71.7; p = 0.095), but not overall survival (p = 0.14). Conclusions: EGFR overexpression is associated with increased tumor aggressiveness as indicated by poor tumor cell differentiation and reduced DFS in colon cancer patients receiving adjuvant 5-FU and levamisole. These findings indicate that EGFR may be an important therapeutic target in the adjuvant setting.

[Table: see text]
A quantitative reverse transcriptase polymerase chain reaction assay to identify metastatic carcinoma tissue of origin
Authors: Talantov D. , Baden J. , Jatkoe T. , Yu J. , Atkins D. , Mazumder A. , Wang Y.
Published: March 2020

20024

Background: Carcinoma of unknown primary (CUP) wherein metastatic disease presents without an identifiable primary tumor site represents approximately 3–5% of all cancers. Identifying the origin of the primary tumor in patients with CUP can enable rational choice of therapeutic regimens. We developed an optimized set of ten gene markers for a quantitative reverse transcriptase polymerase chain reaction (qRTPCR) assay, and demonstrated high accuracy in predicting the tissue of origin when used on with formalin-fixed, paraffin-embedded (FFPE) metastatic carcinoma samples. Methods: Twenty-three putative tissue-specific markers for lung, colon, pancreas, breast, prostate and ovarian carcinomas were nominated by querying a gene expression profile database and by performing a literature search. Ten of these marker candidates were then selected based on validation by qRTPCR on 205 FFPE metastatic carcinomas of known tissue origin. Next, we optimized the RNA isolation and qRTPCR methods for these ten markers, and tested the qRTPCR assay on two sets of FFPE metastatic tumors. Results: We applied the 10-gene qRTPCR assay to a set of 260 metastatic tumors of known origin, generating an overall accuracy of 78%. Furthermore we tested an independent set of 48 metastatic samples, including thirty-seven samples where either the tissue of origin was known or which initially presented as CUP but were subsequently resolved. In these 48 samples, our assay demonstrated an accuracy of 76%. Conclusions: Our results suggested that optimized ten-gene markers qRTPCR assay reliably predicts tissue of origin of metastatic carcinomas in FFPE tissues. Such assay can significantly improve the rate of tissue of origin identification for carcinoma of unknown primary.

[Table: see text]
Patients with breast and ovarian cancer generate immune responses to the folate receptor alpha
Authors: Knutson K. L. , Kalli K. R. , Krco C. J. , Goodman K. , Kelemen L. E. , Low P. S. , Hartmann L. C.
Published: March 2020

20043

Background: Studies have demonstrated that the generation of endogenous immunity to specific tumor antigens is associated with improved prognosis for many cancers. Thus, there has been increased efforts to identify tumor-associated antigens to which there is immunity, as these may be tumor rejection antigens. A candidate antigen is the folate receptor alpha (FRα) which is overexpressed in malignancies, especially breast and ovarian cancer. Prior studies involving ovarian and breast cancer patients have demonstrated the presence of FRα-specific cytotoxic T cells in the tumor-associated lymphocyte population. Methods: Using the RANKPEP CD4 T cell epitope prediction algorithm, we predicted promiscuous immunogenic regions of FRα, and tested for immunity in 30 breast or ovarian cancer patients and 18 healthy volunteer donors in order to attain a better understanding of the levels and extent of the endogenous FRα immune response. Immunity was examined using IFN-γ ELIspot analysis and IgG ELISAs. Results: Fourteen peptides were predicted as potential epitopes to which T cells may respond, seven each from the carboxy- and amino-terminus halves of the protein. Greater than 70% of patients demonstrated T cell immunity to at least one of the fourteen peptides. Patients responded to an average of 3 ± 0.7 peptides while healthy donors responded to only 1 ± 0.5 peptides (p = 0.02). Five peptides were recognized by >25% of patients. Responses to three of these peptides were higher (p > 0.05) in the patients than in healthy donors, suggesting that patients generated immunity upon cancer development. Compared to healthy individuals, patients developed higher immunity to the amino-terminus half of the receptor, suggesting natural epitope spreading (p = 0.03). There was no difference between patients and healthy donors in the immune responses to either non-specific stimuli (p = 0.2) or viral peptides (p = 0.5). Patients had detectable FRα-specific IgG antibody consistent with active FRα-specific helper T cell immunity. Conclusions: These findings demonstrate that the FRα is a target of the immune system in patients with breast or ovarian cancer. Understanding the antigens that are naturally targeted by the immune system may be important for diagnosis, prognosis, and immune-based therapies.

No significant financial relationships to disclose.
Incidence of circulating tumor cells (CTCs) in peripheral blood of breast cancer patients at primary diagnosis—A potential tool for risk stratification
Authors: Rack B. K. , Schindlbeck C. , Janni W. , Jeschke U. , Hofmann S. , Beckmann M. W. , Pantel K. , Lichtenegger W. , Sommer H. , Friese K.
Published: March 2020

20053

Background: In metastatic breast cancer, CTCs have been proven an independent predictor for progression-free and overall survival (Cristofanilli, NEJM 2004). Therefore, we evaluated the role of CTCs in peripheral blood of breast cancer patients at primary diagnosis and during adjuvant systemic treatment within the Success-Trial (n = 3,658 pts). First results of the incidence of CTCs prior to chemotherapy will be presented here. Methods: The CellSearchSystem (Veridex) was used for isolation and enumeration of CTCs from 23 ml of peripheral blood. Briefly, after immunomagnetic enrichment with an anti-Epcam-antibody, cells were labelled with anti-cytokeratin (8,18,19) and anti-CD45 antibodies to distinguish epithelial cells and leukocytes. Patients had been accrued between September and December 2005. Results: We analyzed 124 breast cancer patients at the time of primary diagnosis. The majority of patients presented with small tumors (41% T1, 55% T2, 4% T3), and 32% were node-negative (45% N1, 14% N2, 9% N3). While a positive hormone receptor status was observed in 74% of patients and 31% were her2/neu positive, most tumors showed an intermediate or unfavourable grading (2% G1, 98% G2–3). In this patient group, 32% of patients (n = 40) presented with ≥1CTC in peripheral blood. In patients with the detection of CTCs, the mean number of cells was 5 (range 1–38). In 11 healthy individuals we found 1 CTC in one blood sample. The presence of CTCs did not correlated with tumor size (p = .39), presence of lymph node metastases (p = .25), histopathological grading (p = .33), hormone receptor status (p = .56) or her2/neu status of the primary tumor (p = .14). Conclusions: The isolation of CTCs from peripheral blood of primary breast cancer patients is feasible and might prove as an easy applicable and convenient technique to give further insight into the spread of the disease. The Success-Trial will investigate, whether CTCs can be used for risk stratification to more tailored treatment approaches and for the monitoring of treatment efficacy in individual patients.

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The importance of vascular invasion (vi) in women with breast cancer (bc) and between 1 and 3 positive nodes: Analysis of 272 patients
Authors: Fiorio E. , Auriemma A. , Mandarà M. , Caldara A. , Mercanti A. , Pavarana M. , Micciolo R. , Cetto G. , Molino A.
Published: March 2020

20062

Background: Node positivity and the number of positive nodes are the most important prognostic factors in BC, and affected patients usually have a worse OS. The presence of VI in primary BC is a prognostic factor correlated with a high risk of metastatic spread whose importance led to it being included in the new classification proposed at the VII St. Gallen Consensus Conference. Methods: We studied VI in 272 pts with invasive BC and 1–3 positive nodes in order to evaluate its importance, and correlate it with the pathological, biological and clinical factors of T (1,2,3,4), diameter (0–10 mm, 11–20 mm, 21–50 mm, ≥50 mm), grading (G1, G2, G3), ER and PgR (positive vs negative), Ki-67 proliferative index (low 0–10%; intermediate 11–25%; high 26–100%), c-erbB2 (positive vs negative), age at diagnosis (≤49, 50–65, ≥65 years), type of diagnosis (asymptomatic vs symptomatic) and type of surgery (conservative vs mastectomy). The data were analysed using the chi-squared test. Results: Unlike those without VI, the tumours with node involvement (N+ 1–3) and VI correlated with poor prognostic factors: they were larger (pT2: 62.5% vs 37.5%; p = 0.001), had higher grades (G2: 50.7% vs 49.3%; G3: 72.2% vs 27.8%; p = 0.0004), and were more likely to be ER- (75.6% vs 24.4%; p = 0.004) and c-erbB2+ (50.6% vs 49.4%; p = 0.06). The patients were younger at diagnosis (63% vs 37% aged ≤49 years; p = 0.01) and more likely to have a symptomatic diagnosis (61% vs 39%; p = 0.001). No association with PgR was found. Conclusion: Our results show that primary BC with N+ 1–3 and VI+ correlates with poorer prognostic factors (diameter >20 mm, G2/G3, ER-, c-erbB2+, younger age and symptomatic diagnosis). These correlations are in accordance with the new classification proposed at the VII St. Gallen Consensus Conference concerning high-risk pts with early BC. As these pts are at high risk of recurrence after good local surgery, it is important to offer them adequate adjuvant therapy as a reasonable treatment opinion on the basis of their endocrinal responsiveness.

No significant financial relationships to disclose.
Early placenta insuline-like peptide (pro-EPIL): A novel biomarker in advanced and metastatic non-small cell lung cancer (NSCLC) patients treated by chemotherapy
Authors: Ayllon J. , Oudard S. , Banu E. , Janneau J. L. , Helley D. , Richon S. , Banu A. , Levy E. , Medioni J. , Bellet D.
Published: March 2020

20069

Background: Some studies suggest a causal relationship between the early placenta insuline-like peptide encoded by the insulin-like 4 gene and cancer invasiveness. Methods: Serum concentrations of pro-EPIL were measured by ELISA using a novel two-site “sandwitch” assay, based on two monoclonal antibodies (mAb) raised against synthetic peptides analogous to two distinct regions of the pro-Epil polypeptide: mAb EPIL15, and biotinylated mAb EPIL02, used as the labelled indicator. The first mAb EPIL15 was purified and bound to a solid phase. Pro-EPIL was allowed to bind, and unbound proteins were removed by washing. The sensitivity limit of the assay was 3 ng/mL of synthetic peptide, and detection was linear over a range of at 3 pg to 200 ng of the synthetic peptide. As primary endpoint, we studied the Pro-EPIL distribution according to some clinico-biological prognostic factors. Results: Between July 2001 and April 2005, nineteen chemonaive NSCLC pts were treated by chemotherapy in our center and serum determinations of pro-EPIL were performed. Ten pts (63%) overexpressed pro-EPIL before start of chemotherapy with a median value of 1.06 ng/mL (range 0.01–6.75). Median age was 60 years (range 38–80), more than 90% of pts were men with an advanced or metastatic disease. Forty-two percent of pts had a large cell carcinoma subtype, with 25% of adenocarcinoma and 25% of epidermoid type. Immunologic evaluations were performed with a median value of the CD3+ lymphocites of 1290/mm3 (range 610–2138). Significant correlations were observed between age, serum alcaline phosphatase and CD3+ number. A border-line positive correlation was observed between pro-EPIL and CD3+ levels (P = 0.09, R2 = 0.5). Median OS for entire cohort was 7.5 months (95% CI, 5.5–9.4), with 67% deaths. Because our sample size was very low, no survival analysis were performed according to the baseline pro-EPIL value and other prognostic factors. A bootstrapping procedure is planned on our data. Conclusions: This results showed that pro-EPIL was overexpressed in the majority of advanced NSCLC pts. Interactions between this biomarker and immune system are possible. Furthemore, pro-EPIL as a therapeutic target might be tested in prospective studies.

No significant financial relationships to disclose.
Clinical and biologic relevance of interferon (IFN) regulated proteins in esophageal squamous cell carcinoma
Authors: Guimaraes D. P. , Oliveira I. M. , Paiva G. R. , Souza D. M. , Marques C. B. , Olmedo D. B. , Pinto C. E. , Hainaut P. , Small I. , Ferreira C. G.
Published: March 2020

20077

Background: The prognosis for SCCE is poor, with less than 10 percent of patients surviving beyond five years. The ability to identify specific markers associated with biological behavior may provide information about the likelihood of both localized and locally advanced disease. The Interferon regulatory factor 1 (IRF-1) act as a tumor suppressor protein by affecting cell cycle progression and/or apoptosis. The Guanylate binding protein-2 (GBP-2) is an interferon regulated protein whose induction requires the IRF-1 activation. Methods: We analyzed the IRF-1 and GBP-2 expression by IHC in 42 patients (pts) with surgically ressected SCCE. A detailed pathologic examination was conducted by selecting single histologic section, which contained a clear transition zone from normal epithelium to invasive cancer. Staining was semiquantitatively scored by two observers based on both intensity and percentage of stained cells. Clinical data information was obtained from medical records. Results: The median age was 58 (39–75); Male/female 61.9/38.1%; Of 39 pts, 53,8% had localized disease (I/IIA) and 46,2% had locally advanced disease (IIB/III). Nineteen pts received standardized radiochemotherapy before surgery. The IRF-1 and GBP-2 were expressed in 100% and 75% of tumors, respectively. Normal epithelium was stained for both IRF-1 and GBP-2 with distinct expression patterns. There was a significant correlation between scores for GBP-2 and IRF-1 tumor staining (p = 0.23). Interestingly, the score for GBP-2 tumor staining was inversely correlated to clinical stage (p = 0.001), tumor invasion of the esophageal wall (T) (p = 0.26), involvement of lymph nodes (N) (p = 0.035) and tumor extension (p = 0.007). There was no correlation between the score of IRF-1 staining and the clinical-morphological parameters. Conclusions: We reported for the first time the expression of GBP-2 protein in esophageal cancer. Our results suggest that GBP-2 expression is related to less aggressive behavior of SCCE. Further studies are required to clarify biological GBP-2 protein function, and possible prognostic implications.

No significant financial relationships to disclose.
MBD4 and SEI1 as new prognostic markers in colorectal cancer patients receiving adjuvant 5-fluorouracil
Authors: Ju J. , Formentini A. , Nakajima G. , Kornmann M. , Xi Y.
Published: March 2020

20085

Background: Previous studies from our laboratory have identified a number of marker genes associated with chemosensitivity to 5-fluorouracil (5-FU). In this study, the in vivo significance of several marker genes in terms of prognostic potential were evaluated using colorectal cancer patient samples treated with 5-FU based therapy. Eight marker genes were selected based on their functional roles and significant fold changes in expression. They are SERTA domain containing 1(SEI1), Ribonucleotide reductase M2 polypeptide (RRM2), Origin recognition complex, subunit 6 homolog-like (ORC6L), Eukaryotic translation initiation factor 4E (EIF4E), Thymidylate Synthase (TS), SET and MYND domain containing 3 (SMYD3), Dickkopf homolog 4 (DKK4) and Methyl-CpG binding domain protein 4 (MBD4). Methods: Forty eight snap frozen clinical colorectal samples (24 normal and 24 paired colorectal cancer patient samples) were selected with detailed clinical follow-up information. cDNAs were synthesized and the expression levels of marker genes were quantified via qRT-PCR analysis. The statistical significance of these markers for disease prognosis was evaluated using two tailed paired T-test. Kaplan-Meier survival curve was generated and analyzed by Logrank test. Results: Based on the quantitative expression analysis, RRM2 (p = 0.0001, 95% CI, 2.0 to 4.5), ORC6L (p = 0.0001, 95% CI, 1.8 to 4.6), EIF4E (p = 0.0002, 95% CI, 0.3 to 0.9), TS (p = 0.0005, 95% CI, 0.7 to 2.2) and SMYD3 (p = 0.0001, 95% CI, 0.8 to 1.5) was over-expressed in tumor tissues. However, the expression of SEI1 was decreased in tumors (p = 0.02, 95% CI, 0.1 to 1.3), consistent with the function of SEI1 as a potential tumor suppressor. Kaplan-Meier survival analysis indicated that MBD4 is a significant prognostic factor for patient’s survival (p = 0.03). MBD4 was a key protein involved in DNA methylation. The expression of TS was associated with tumor stage as it had significant higher expression level in UICC stage I and II compared to stage IV patients (p = 0.03). Conclusion: MBD4 may be potential novel prognostic markers for predicting patient survival for colorectal cancer.

No significant financial relationships to disclose.
registHER: Baseline characteristics of a cohort of HER2-positive metastatic breast cancer (MBC) patients
Authors: Kaufman P. , Mayer M. , Paik S. , Ulcickas Yood M. , Yardley D. , Tan-Chiu E. , Brufsky A. M. , Rugo H. , Tripathy D. , Wang L.
Published: March 2020

20095

Background: HER2 is amplified in 25% of breast cancers and is associated with poor survival. registHER captures the natural history, treatment patterns and outcomes in 1000 newly-diagnosed HER2-positive MBC patients (pts) throughout the U.S. This observational study recruits pts in both academic and community centers. Methods: This ongoing prospective cohort study collects clinical, pathologic and treatment data at enrollment, quarterly until death, loss to follow-up or 3 years after the last enrollment. We describe baseline pt and clinical characteristics in registHER compared with HER2-positive MBC pts in the phase III pivotal trial (Slamon DJ, et al. N Engl J Med. 2001;344:783–792). Results: Between December 2003 and September 2005, 813 eligible pts were enrolled at 280 study sites. Most pts were seen at community-based (76%) vs academic (18%) clinics; a few pts did not fall into either category (6%). A comparison of baseline characteristics is shown below. Conclusions: registHER pts tended to have a shorter disease-free interval and more estrogen receptor positive disease than pts in the pivotal trial. Reasons for these differences could reflect trial referral and/or diagnostic testing differences. Fewer registHER patients were white, but other characteristics were similar between the two groups. These findings support the hypothesis that observational studies describe a broad patient population which may not exactly duplicate clinical trials. Within registHER, there was some variation between academic vs community clinics (eg. nodal status and adjuvant therapy). Treatment pattern analyses are ongoing.

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