bluuebunny/crossref_metadata_2025 huggingface dataset:
Perspective evaluation of emotional state of oncological patients in the waiting rooms
Authors: La Verde N. M. , Resega R. , Lurati C. , Garassino M. C. , Torri V. , Cinquini M. , Buffa G. , Landi Mrs J. , Riva M. , Borgonovo K. , Farina G.
Published: August 2020

e20693

Background: The waiting room is one of the most emotional place in oncological divisions; indeed, it's presumed that the waiting moment makes increase thoughts most related to the experience of disease and its treatment, making them sometimes unbearable. At present, any evaluation of the emotional states in the waiting room doesn't exist, therefore we analyzed the most prevailing emotions and feelings for patients in this context. The identification of them should address also ways to improve patients well being in these moments. Primary objective was to prospectively evaluate the prevailing emotional state patients and feelings in oncological waiting rooms; secondary endpoint is to develop a questionnaire to be further validated and useful to quantify how changes in the environment, as people and furnishings, could reduce prevailing emotional intensity. Methods: A committee of psychoncologists, oncologists, psychiatrists, volunteers, nurses, past patients, and statisticians identified the emotional states considered most important (relationship with environment, surrounding people and treatment, recurrent thoughts about disease and life, fears and things that could ease the wait and prevailing emotions) and developed a questionnaire. In May 2008 for two consecutive weeks this questionnaire was administered by the same dedicated nurse to all the outpatients afferred to our Oncology Department. The compilation of questionnaire was spontaneous and anonymous and it collected also age, sex, whether the patient was under treatment or in follow up and whether was accompanied. Results: In two consecutive weeks 300 questionnaires have been collected in the waiting rooms. Statistical analysis showed that the most prevailing emotions are sadness (51%), fear (29%), anger (17%) and joy (7%). A positive disposition (Cronbach alpha 0.60) and anxiety (Cronbach alpha 0.64) were also assessed. No differences in answers were revealed among patients waiting for follow up and patients waiting for chemotherapy and among different ages. Conclusions: The level of negative feelings seems higher than in other conditions and suggests that active intervention to improve the waiting in waiting rooms is warranted. A further validated questionnaire is ongoing also to evaluate emotions intensity.

No significant financial relationships to disclose.
Assessment of quality of life using FACT-G survey in a phase I trial in cancer patients
Authors: Jung G. , Knight D. , Moadel A. , Desai K. , Chaudhary I. , Gajavelli S. , Ghalib M. H. , Mani S. , Goel S.
Published: August 2020

e20709

Background: Quality of life (QoL) assessment in clinical trials has been gaining more attention. FACT-G surveys have been validated to assess QoL in clinical trials involving oncology patient (Cella DF et al, J Clin Oncol 11:570–579, 1993). However, there is paucity of evaluation of QoL in patients with advanced cancer participating in Phase I clinical trials. Methods: FACT-G surveys were conducted within the context of a Phase I trial to identify a safe dose and potential drug-drug interations of capecitabine and irinotecan combination (Goel, S et al, Invest New Drugs 25:237–245, 2007). The FACT-G survey consists of 28 questions in 5 sections, namely, physical well-being, social/family well-being, emotional well-being, relationship with doctor, and functional well-being). Patients were requested to complete the FACT-G surveys at baseline and every two cycles thereafter (each cycle of 3 weeks duration). Results: Forty-one of 47 patients with advanced solid tumors who participated in the clinical trial completed FACT-G surveys. Mean scores were calculated for each time point. The mean QoL scores at baseline and post cycle 2 were 53 and 58, respectively (p = 0.1). Post cycle 4, the mean QoL score was 62 [p = 0.01, (vs. baseline)]. Following cycle 4, the number of respondents decreased to the extent where we were unable to ascertain any further changes in the QoL scores. Conclusions: It is feasible to use FACT-G survey as a tool to assess QoL in patients participating in an oncology phase I clinical trial. Although the sample size of the patient population was not powered for any statistical significance, there was a trend toward improving QoL based on FACT-G survey scores. This suggests that phase I clinical trials may provide improvement of QoL for some patients. FACT-G is a useful tool in assessing QoL in oncology phase I trial study population.

No significant financial relationships to disclose.
Evaluation of side effects in cancer patients during oncological care: A project of the supportive care group of the tumor center Berlin
Authors: Feyer P. C. , Steiner U. , Bangemann N. , Kurz S. , Rudolph C. , Schelenz C. , Jagota A.
Published: August 2020

e20715

Background: The therapeutic options in tumour treatment protocols include a maximum of tumour cell reduction and at the same time a minimum of side effects in order to secure a good quality of life for the cancer patient. Temporary side effects have been accepted in relation to the benefit risk calculation. A questionnaire for cancer patients was developed to improve the management of side effects of the cancer treatment, to optimize the patient-doctor communication and to register individual problems of the patient. Methods: Cancer patients received a questionnaire with 25 items and were asked for their problems during or after the last tumour specific treatment before having the consultation with the doctor. Gender, age, Karnofsky-index and treatment protocol were additional registered. During September 2007 until December 2007 a total of 272 questionnaires could be collected. The qualitative items have been evaluated with the contigens table method and the parameter free x2 test (Chi-Square-Test). Results: More than 50% of the patients documented additional important points influencing their well being. Nearly all of the evaluated patients had some tumour therapy specific side effects. The most pronounced problem was the physical exhaustion in 71% of the evaluated patients. More than 50% were influenced in their quality of life by more than normal fatigue since the start of their tumour specific therapy. Significant differences could be evaluated in relation to the age, diagnosis and tumour therapy with respect to the symptoms. Conclusions: The questionnaire was be found to be a good method to improve the communication between the patient and the doctor. It was easier to point out individual problems for the patient. The care givers can focus more precisely on important side effects of the tumour specific therapy in order to improve the quality of life of the treated patient.

No significant financial relationships to disclose.
Assessment of efficacy, safety, and tolerability of aprepitant in adult patients with chemotherapy-induced vomiting
Authors: Sharma A. D. , Julka P. K. , Rastogi A. , Suri T. , Rath G. K. , Sharma B. N. , Baliga V. P.
Published: August 2020

e20729

Background: Approximately 70 to 80% of all cancer patients receiving chemotherapy experience emesis and 10 to 44% experience anticipatory emesis. Aprepitant a neurokinin 1 (NK 1) receptor antagonist is used in chemotherapy induced nausea and vomiting. The present study was undertaken to assess efficacy, safety, and tolerability of Aprepitant in adult patients with chemotherapy induced nausea and vomiting. Methods: The study was conducted as a prospective, multicenter, randomized, open label, single group, phase III study in adult patients with chemotherapy induced nausea and vomiting and patients admitted to hospital for any type of cancer. The final protocol was approved by the Institutional ethics committee before the initiation of the study. Informed consent was obtained from all the patients before enrollment in the study. Aprepitant capsules were given in the dose of 125mg orally one hour prior to chemotherapy on day 1 followed by 80 mg orally in the morning on days 2 & 3.Aprepitant was combined with oral Dexamethasone 12mg oral 30 minutes prior to chemotherapy on day one and 8mg on the following two days. Efficacy of aprepitant was evaluated through number of emetic episodes within 24 hours or requirement for the rescue medication. Self assessment of nausea was done by patients using a 100 mm visual analog scale. Safety and tolerability was assessed through monitoring of adverse events and physical examination. Results: A total 30 patients (13 males) (mean age 40.45 years) were enrolled in the study. Significant (p<0.05)changes in emetic episodes were seen after treatment with aprepitant. Overall global assessment of vomiting indicated that 19 patients (82.6 %) had very good improvement, 3 patients (13%) had good improvement and one patient (4.4%) showed fair improvement after treatment with aprepitant. The most common adverse events included fever, headache, diarrhoea skin rash, constipation, and blisters. Severity of adverse events was mild to moderate and the adverse events disappeared with continued therapy. Conclusions: Treatment with aprepitant is a useful therapeutic option for the management of chemotherapy induced nausea and vomiting.

[Table: see text]
Cognitive disfunctions in women with breast cancer treated with adjuvant chemotherapy: A preliminary report
Authors: Gercovich D. , Hirsch H. , Lopez P. , Gercovich N. , Vázquez L. , Gil Deza E. , Martin Reina G. , Margiolakis P. , Morgenfeld E. , Gercovich F. G.
Published: August 2020

e20742

Background: Recent studies suggest that women diagnosed with breast cancer and treated with adjuvant chemotherapy may suffer cognitive deterioration. These studies, however, present a number of limitations including a small sample of patients, retrospective evaluation of cognitive skills, lack of cognitive-function measurements previous to the chemotherapy and a lack of evaluation of psycho-pathological factors that could potentially influence on the cognitive function of the subjects. The aim of this study is to present the preliminary results of a prospective study about the impact of conventional adjuvant chemotherapy on a series of cognitive domains in adult women diagnosed with breast carcinoma, monitoring frequent psycho-pathological variables such as anxiety and depression. Methods: The present is a cohort, observational study. It includes patients with a diagnosis of stage I and II breast carcinoma, treated with standard chemotherapy and able to give their consent. The protocol has 3 stages of evaluation. First evaluation: a basal measurement between diagnosis and the first chemotherapy course. The second and third evaluations are done at 8 and 14 months respectively after the first one. Each patient has been studied using a neuro-psychological battery of 17 different tests which evaluate the following cognitive domains: global cognitive functioning, intelligence, memory, language and executive functions. The levels of anxiety and depression were studied simultaneously. Further tests will be done for related samples, so as to compare changes within a group. Results: Up to date 14, patients have completed the basal battery of tests. We expect to find differences in the cognitive functions of the patients treated with adjuvant chemotherapy. The aforementioned differences will be controlled with psycho-pathological measurements. Conclusions: The present study will allow us to evaluate in a prospective and exhaustive way cognitive deficiencies in women with breast carcinoma treated with adjuvant chemotherapy. In addition, the use of psycho-pathological instruments will allow monitoring and understanding the neuro-psychological results in a more reliable way.

No significant financial relationships to disclose.
Amplification of RUNX1 gene in two new cases of childhood B-cell precursor acute lymphoblastic leukemia: A case report
Authors: Fauzdar A. , Mahajan A. , Jain D. , Mishra M. , Raina V.
Published: August 2020

e21000

Background: Chromosome abnormalities of leukemia cells have important prognostic significance in childhood acute lymphoblastic leukemia (ALL). B-cell precursor acute lymphoblastic leukemia (BCP-ALL) ETV6/RUNX1 (alias TEL/AML1) is most frequent i.e. 15 - 35% in the children with 2 - 18 age group. We report two new cases with Pre B- cell ALL without ETV6/RUNX1 rearrangement, showing amplification of AML1 gene detected by FISH analysis. Methods: Bone marrow samples were analyzed for chromosomal abnormalities with conventional G-banding techniques and interphase fluorescence in situ hybridization (FISH) using probes to detect BCR/ABL t(9;22)(q34-q11) fusion, cryptic TEL/AML1 t(12:21)(p13-q22) and MLL rearrangement for del 11q23. Results: In first case a 3-year girl with four copies of AML (RUNX1) gene were observed in 95% of the cell with normal two copies of TEL (ETV6) gene in both interphase and metaphase FISH. We observed BCR-ABL negative translocation and no MLL gene rearrangement in all the interphase cells after doing FISH. We got a normal 46XX karyotype from bone marrow with conventional cytogenetics (CC) in the same patient. In second case, a 4-year male we observed four copies of AML and two copies of TEL gene in more than 80% of cells. In this patient, we got BCR-ABL negative translocation and three copies of MLL gene without any rearrangement through FISH. We got normal 46XY karyotype in the same patient through CC. Conclusions: In both the patients, we observed hyperdiploidy detected with four copies of RUNX1 gene showing tetrasomy of chromosome 21 detected with metaphase FISH analysis whereas G-banding shows normal diploidy. Bone marrow karyotype in combination with molecular cytogenetic techniques like FISH should be done for improvement in sensitivity and accurate cytogenetic analysis in childhood ALL patients for proper identification of prognostic group for optimum treatment. This is one of the few reported studies worldwide for amplification of RUNX1 gene from Indian subcontinent in childhood BCP-ALL.

No significant financial relationships to disclose.
Ewing tumors of the chest wall: Local control and long-term outcomes
Authors: Indelicato D. J. , Keole S. R. , Shahlaee A. H. , Morris C. G. , Gibbs C. P. , Scarborough M. T. , Islam S. , Marcus R. B.
Published: August 2020

e21501

Background: Primary sarcomas of the lungs and mediastinum are rare and few data are reported on treatment and results of therapy. Methods: We reviewed our experience from 1980 and 2008 including 31 patients (pts). Pts characteristics: median age 41 (19–80 y), male/female 19/12; symptoms at diagnosis: dyspnoea (42%), chest and shoulder pain (39%), cough (35%), hemophtoae (13%), discomfort (10%). 4 pts had a previous history of mediastinal radiation for Hodgkin's and non-Hodgkin's linfomas. 5 mediastinal tumours were located as follows: 2 in anterior part, 1 in posterior and 2 in the middle (sarcomas of the heart). 26 lung sarcomas presented as a singular mass in 23 cases and as a metastatic disease in 3. Results: In 20/31 cases the tumour was immediately resected (3 mediastinal masses and 17 lung sarcomas). In 8/31 cases only biopsy was possible. FNA was done in 25 pts. Neoadjuvant chemotherapy was performed in 4 cases (3 resected). Resection was complete in 11/23 cases and in 12/23 incomplete. The histology were: peripheral nerve tumour 7, leiomyosarcoma 4, MFH 2, fibrosarcoma 2, liposarcoma 1, angiosarcoma 2, undifferentiated sarcoma 1, solitary fibrous tumour 2, rhabdomyosarcoma 2, synovialsarcoma 2, pulmonary artery sarcoma 1, pleuropolmonary blastoma 1, malignant hemangiopericytoma 1, mixoid chondrosarcoma 1, ectopic osteosarcoma 1, aggressive fibromatosis 1. Only 4 pts received neoadjuvant chemotherapy, 11 adjuvant CT, 5 exclusive CT + RT for inoperable disease. Radiotherapy was completed in 26 pts (21 adjuvant). Local relapse or metastatic progression were recorded in 16/23 pts and 12 received one or more lines of palliative CT. Data about survival are disposable only for the more recently recorded pts (1998–2008: 17 pts). Of these only 8 are alive (2 with disease). Volume of disease, complete resection and grading are the dominant prognostic factors. Conclusions: Primary sarcomas of the lungs and mediastinum have a very severe prognosis. Surgical resection is the fundamental therapy, but in the future the role of neoadjuvant CT will increase.

No significant financial relationships to disclose.
Ezrin immunohistochemical expression in advanced soft tissue sarcoma biopsy specimens from patients treated in a phase II trial of dose-dense doxorubicin- and ifosfamide-based chemotherapy
Authors: Almeida G. F. , Siqueira S. A. , Castro G. , Snitcovsky I. L. , Akaishi E. H. , Camargo O. P. , Oliveira C. R. , Filippi R. Z. , Federico M. H.
Published: August 2020

e21508

Background: Ezrin is a member of the ERM (ezrin, radixin, moesin) protein family and links F-actin to the cell membrane, and it is involved in regulating growth and metastatic behaviour of cancer cells. Ezrin expression is associated with tumor progression and metastasis in several cancers, including sarcomas. This study evaluated ezrin expression as a predictor of response to chemotherapy and as a prognostic factor in soft tissue sarcoma (STS) patients (pts) treated in a dose-dense schedule protocol. Methods: 21 chemotherapy-naïve pts diagnosed with high-grade STS, not candidates for a limb-sparing surgery, were enrolled in a prospective phase II study of a sequential and dose-dense regimen consisting of doxorubicin 30 mg/m2 d1–3 q2w and ifosfamide 2.5 g/m2 d1–5 q3w, 3 cycles each, with G-CSF support. Ezrin expression was analyzed by immunohistochemistry on slides from formalin-fixed, paraffin-embedded, primary tumor biopsy blocks, with the anti-ezrin antibody clone AB-1(3C12) (NeoMarkers). Cytoplasmic immunostaining in more than 10% of tumor cells was considered as positive. Ezrin expression was correlated with response rate, progression-free (PFS) and overall survival (OS). Results: Leiomyo-, synovial and sarcoma NOS were the most frequent subtypes (5 pts each). 13 out of 21 pts (62%) presented distant metastasis. Protocol was halted after 3 toxic deaths. Three pts achieved partial response (RR 14%). With a median follow-up of 11.7 mo, median PFS and OS were 8.9 and 20.1 mo, respectively. In univariate analysis, OS was higher for synovial sarcoma pts (not reached vs. 14.2 mo, HR 0.0, 95%CI 0.06–0.82, p=0.02), and for those aged 45 y or less (20.1 vs. 4.2 mo, HR 0.30, 95%CI 0.03–0.88, p=0.04). Ezrin expression was available for 20 pts: it was positive in 9 pts (45%) and negative in 11 pts (55%). Ezrin expression was not related to tumor response (p = 0.40), and no significant association was detected between ezrin expression and median PFS or OS. Conclusions: Expression of ezrin was not a useful marker to predict outcomes in STS pts treated with dose-dense doxorubicin- and ifosfamide-based chemotherapy.

No significant financial relationships to disclose.
Primary sarcomas of the lungs and mediastinum: Clinicopathological study and therapy results of Piedmontese Group for Sarcomas
Authors: Comandone A. , Boglione A. , Pochettino P. , Berno E. , Inguì M. , Papotti M. , Borasio P. , Maggi G. , Brach del Prever E. , Gino G.
Published: August 2020

e21509

Background: Primary sarcomas of the lungs and mediastinum are rare and few data are reported on treatment and results of therapy. Methods: We reviewed our experience from 1980 and 2008 including 31 patients (pts). Pts characteristics: median age 41 (19–80 y), male/female 19/12; symptoms at diagnosis: dyspnoea (42%), chest and shoulder pain (39%), cough (35%), hemophtoae (13%), discomfort (10%). 4 pts had a previous history of mediastinal radiation for Hodgkin's and non-Hodgkin's linfomas. 5 mediastinal tumours were located as follows: 2 in anterior part, 1 in posterior and 2 in the middle (sarcomas of the heart). 26 lung sarcomas presented as a singular mass in 23 cases and as a metastatic disease in 3. Results: In 20/31 cases the tumour was immediately resected (3 mediastinal masses and 17 lung sarcomas). In 8/31 cases only biopsy was possible. FNA was done in 25 pts. Neoadjuvant chemotherapy was performed in 4 cases (3 resected). Resection was complete in 11/23 cases and in 12/23 incomplete. The histology were: peripheral nerve tumour 7, leiomyosarcoma 4, MFH 2, fibrosarcoma 2, liposarcoma 1, angiosarcoma 2, undifferentiated sarcoma 1, solitary fibrous tumour 2, rhabdomyosarcoma 2, synovialsarcoma 2, pulmonary artery sarcoma 1, pleuropolmonary blastoma 1, malignant hemangiopericytoma 1, mixoid chondrosarcoma 1, ectopic osteosarcoma 1, aggressive fibromatosis 1. Only 4 pts received neoadjuvant chemotherapy, 11 adjuvant CT, 5 exclusive CT + RT for inoperable disease. Radiotherapy was completed in 26 pts (21 adjuvant). Local relapse or metastatic progression were recorded in 16/23 pts and 12 received one or more lines of palliative CT. Data about survival are disposable only for the more recently recorded pts (1998–2008: 17 pts). Of these only 8 are alive (2 with disease). Volume of disease, complete resection and grading are the dominant prognostic factors. Conclusions: Primary sarcomas of the lungs and mediastinum have a very severe prognosis. Surgical resection is the fundamental therapy, but in the future the role of neoadjuvant CT will increase.

No significant financial relationships to disclose.
Mechanisms of nicotine-induced resistance to treatment with chemotherapy and radiotherapy
Authors: Warren G. W. , Kudrimoti M. , McGarry R. , Arnold S. , Rangnekar V.
Published: August 2020

e22008

Background: Nicotine (NIC), used for smoking cessation in cancer patients, is associated with increased tumor growth, angiogenesis, migration, invasion, growth factor production, and inhibition of apoptosis. The effects of NIC on the efficacy of radiotherapy (XRT) and/or chemotherapy (CT) have not been well characterized. Methods: A review of the literature produced a construct demonstrating that NIC generates more than 100 interactions with 40 proteins involved in tumor promotion and progression. From this construct, nicotinic acetylcholine receptors (nAChR), PI3K/Akt, and MAPK were identified as potential critical upstream targets modulating the effects of NIC. The effect of NIC on cell proliferation, cell survival, and protein activation was analyzed following CT, XRT, or chemoradiotherapy (CRT) in FaDu, A549, and H460 human cancer cell lines. Results: Proliferation measured with the WST-1 (tetrazolium) assay demonstrated that NIC increased proliferation following cisplatin, XRT, and cisplatin with XRT. NIC administration increased cell survival (colony survival) following XRT and following CT (cisplatin, doxorubicin, or etoposide) regimens in all cell lines. nAChR expression was verified in each cell line using Western blot. Non-specific inhibition of nAChR demonstrated no consistent effect on preventing NIC induced resistance to CT, XRT, or CRT; however, specific inhibition of the alpha-3- or alpha-7-nAChR had variable cell specific ability to prevent NIC induced resistance. Inhibition of MAPK activity (using PD98059) produced marked reduction in proliferation of all cell lines, but generally had no effect on NIC induced resistance. PI3K inhibition (using LY294002) generally increased sensitivity to cisplatin and XRT; however, some PI3K activity was required to achieve the full effect of cisplatin. NIC prevented LY294002 induced sensitization to chemotherapy with no effect on sensitization to XRT. Conclusions: Nicotine induces a broad spectrum of resistance to CT, XRT, and CRT and serves as an excellent model of treatment resistance. Mechanisms of NIC induced resistance to CT and XRT appear to diverge. These findings have significant clinical implications including smoking cessation, treatment efficacy, and outcomes research in cancer patients.

No significant financial relationships to disclose.
Validation of a microarray-based gene expression test for tumors with uncertain origins using formalin-fixed paraffin-embedded (FFPE) specimens
Authors: Pillai R. , Deeter R. , Rigl C. T. , Halks-Miller M. , Henner W. D. , Buturovic L.
Published: August 2020

e22015

Background: Microarray-based gene expression has been validated as an aid in the diagnosis of tumors with uncertain origins when the specimen is frozen tissue. Microarray use has been largely limited to RNA derived from frozen specimens. This study evaluated performance of a microarray-based test in identifying the tumor type in FFPE specimens. Methods: ZFFPE human tumor specimens (n=405) representing the 15 tissue of origin sites on the Pathwork® Tissue of Origin Test panel were blinded and evenly distributed between two independent processing labs. All specimens consisted of a 10-μm-paraffin curl containing at least 60% viable tumor and were either metastatic or poorly differentiated primaries. Each specimen was processed through RNA extraction, amplification, labeling, hybridization to a Pathchip® microarray, and was scanned to generate a qualified data file. A pre-specified classification algorithm utilizing more than 1500 genes was applied to each data file to yield Similarity Scores corresponding to the 15 tissues on the test panel. Results were then unblinded and compared to the available diagnoses. Results: Of the 405 specimens, 352 yielded qualified data files (87%). Based on the top Similarity Score, the overall agreement with available diagnoses was 89% (95% CI, 85%-92%) and for each specimen an average of 12 out of 15 tissues could be ruled out with > 99% probability. Results for all tissue types were highly informative with diagnostic odds ratios ranging from 178 to 28509. Performance was similar for metastatic (n=150; 91% agreement) and poorly differentiated primary specimens (n=202; 87% agreement). Conclusions: The large size of this study allows an accurate estimate of the confidence of test predictions for both ruling in and ruling out tissues as likely sites of primary origin. The Pathwork Tissue of Origin Test makes the potential benefits of microarray-based gene expression tests for tumors with uncertain origins available for use with the most common type of histology specimen, FFPE.

[Table: see text]
Correlation of CXCR4 expression in resected pancreatic adenocarcinoma (PA) with relapse and survival after adjuvant radiochemotherapy (RCT)
Authors: Maréchal R. , Demetter P. , Berton A. , Salmon I. , Van Laethem J.
Published: August 2020

e22022

Background: The chemoreceptors CXCR4, CXCR7 and the hypoxia inductible factor-1 alpha (HIF-1α) are implicated in PA growth, dissemination and angiogenesis. These protagonists are expected to play a key role in radiotherapy and chemotherapy resistance in PA. Methods: We conducted a retrospective study of patients undergoing curative surgery (R0 resection) for PA between 2001 and 2006 in our institution. All were treated with adjuvant RCT (treatment group). The treatment group was case-matched with resected PA patients who did not receive any adjuvant therapy (control group). FFPE specimens were subjected to immunohistochemical analysis using tissue microarray and monoclonal antibodies against human CXCR4, CXCR7, and HIF-1α. Based on the intensity (I) and the extend (E) of staining, cases were stratified into those with high (E x I>3) or low (E x I ≤ 3) expression of CXCR4, CXCR7 and HIF-1α and results were correlated with disease-free survival (DFS) and overall survival (OS). Results: 31 PA patients (median age: 57years, range: 39–76) were analysed in the treatment group and 30 (median age: 59 years, range: 38–81) in the control group. The two groups were well-matched in terms of age, sex, tumor stage (T1-T2 vs T3-T4), tumor differentiation (poor vs well-moderate), lymph node (LN) status (N0 vs N+), lymphatic and vascular embols. In univariate analysis, CXCR4 expression and LN status were associated with DFS and OS (OS, CXCR4low/high: HR, 5.43, 95%CI: 2.03–14.49, p=0.001; N0/N+: HR: 4.62, 95% CI, 1.33–13.12, p=0.016; DFS: CXCR4low/ CXCR4high : HR, 3.01, 95%CI: 1.21–7.46, p=0.018; N0/N+: HR: 3.30, 95% CI, 1.09–9.90, p=0.034) in the treatment group while in the control group LN status was the only variable significantly correlated with DFS and OS. CXCR4 appeared to be the only independant predictor for DFS (CXCR4low/high: HR, 4.95, 95%CI: 1.68–14.71, p=0.007) and OS (CXCR4low/high: HR, 4.76, 95%CI: 2.03–14.49, p=0.004) in the treatment group but not in the control group. Conclusions: CXCR4 was an independant predictor for DFS and OS after adjuvant RCT in resected PA patients. This chemoreceptor could be implicated in the resistance of pancreatic cancer cells to RCT and its targeted inhibition deserves clinical evaluation.

No significant financial relationships to disclose.
Correlation of hENT1 expression with response and survival in non-small cell lung cancer patients treated with gemcitabine-containing chemotherapy
Authors: Oguri T. , Achiwa H. , Ozasa H. , Nakao M. , Uemura T. , Ohta C. , Takakuwa O. , Miyazaki M. , Maeno K. , Sato S. , Ueda R.
Published: August 2020

e22032

Background: The most active gemcitabine uptake has been found via the human equilibrative nucleoside transporter 1 (hENT1). This study was to explore the prognostic impact of the hENT1 on response and survival in Non-small lung cancer (NSCLC) patients treated with gemcitabine-containing chemotherapy. Methods: We developed polyclonal antibody for hENT1. Then we stained hENT1 expression by immunohistochemical analysis in 24 biopsy samples of NSCLC which was formaline-fixed, paraffin- embedded tissues. We were treated with gemcitabine alone or gemcitabine-containing chemotherapy until third-line regimen. Results: They comprised 16 males and 8 females with a median age of 63 years (range 45–82 years). Seventeen patients had adenocarcinomas, six had squamous-cell carcinomas, and one had a large-cell carcinoma. All patients were treated with gemcitabine- containing chemotherapy, with 9, 12, and 3 patients receiving this as a first-, second-, and third-line therapy, respectively. The hENT1-positive staining in NSCLC samples was significantly associated with response to gemcitabine-containing chemotherapy (Fisher's exact test, P<0.05). Responses to gemcitabine-containing chemotherapy were evident in none of the seven patients with no hENT1 expression. Further 3 years survival differed by hENT1 staining: 714 days for hENT1-positive, 316 days for hENT1-negative (HR 2.86; 95%CI 1.13–15.16, P<0.05). Conclusions: While there are some determinants for gemcitabine sensitivity, hENT1 expression may be a predictive maker for the response and survival to gemcitabine-containing chemotherapy in NSCLC.

No significant financial relationships to disclose.
HER2 FISH ratio cut-points and pathologic complete response (pCR), residual tumor (RT), HER2 status, and survival prediction in HER2-positive breast cancer (BC)
Authors: Mehta R. S. , Jackson D. , Schubbert T. , Hsiang D.
Published: August 2020

e22033

Background: We demonstrated that pCR is correlated with increasing HER2-FISH ratio, while disease-free survival (DFS) with pCR and ER-positivity in HER2-positive breast cancer treated with trastuzumab (SABCS 2008). It is known that quantitative HER2-FISH ratio correlates with ER levels and HER2-positivity imparts a higher grade in ER-positive BC. Collectively, we hypothesized that combined ER (≥10) and a HER2 ratio cut-point may subdivide HER2-positive BC into pCR predictive subtypes.Methods: Of the 80 HER2-positive (IHC3+/FISH+) BC, quantitative HER2 FISH ratio (widely spread over 1–18.3) and ER correlation was noted (r=0.34, p=0.002). Moreover, HER2 ratio (>4) correlated with higher Ki-67 (r= 0.5, p=0.01) and grade (p trend=0.05) in ER-positive subtype, inferring a biologic cut-point. Results: Of patients with stage I-IV BC treated neoadjuvantly (92% trastuzumab-based), pCR was 0% (0/13) in ER-positive-low-HER2 compared to 77% (10/13, p=0.0001), 75% (24/32, p<0.0001) and 37.5% (3/8, p=0.043) in ER-positive-high-HER2, ER-negative-high-HER2 and ER-negative-low-HER2, respectively. DFS was 100, 90, 80% and 60% (logrank-trend p<0.05) in these 4 subtypes (excluding stage IV), respectively, at a median follow-up of 38 months (range 6–72). In ER-negative subtypes, DFS was 97% and 29% (logrank p=0.0001) in patients with or without pCR; of the six with RT, 0% DFS was noted in four with HER2-negative/HER2-reduced (HER2-R) RT, compared to 100% in the two with unchanged HER2 (p=<0.05, logrank test). In ER-positive subtypes, DFS is 95% overall, and 100% in patients with RT; 7 of 10 tested RT were HER2- R. Conclusions: pCR is crucial and high in ER-negative-high-HER2 and is crucial (but low) in ER-negative-low-HER2-positive BC for improved outcome. Improved DFS is associated with high pCR in ER-positive-high-HER2 BC, but is independent of the low pCR in ER-positive-low-HER2-subtype. Thus, combined HER2 and ER offer improved prediction. In hypothesis generating analysis, HER2-R may underlie relapse in ER-negative subtypes (HER2-basal-transitional-residual), while it may be beneficial in ER-positive subtypes (luminal-B- A-transitional) by reducing HER2-pathway mediated endocrine resistance.

[Table: see text]
Informative rate and reproducibility of the investigational GeneSearch ProCaM assay in a multicenter laboratory setting
Authors: Baden J. , Markiewicz J. , Painter J. , Jones J. , Curtin K. , Canning S. , Quijano J. , Guinto W. , Wang Y. , Green G.
Published: August 2020

e22038

Background: PSA tests have low specificity, which frequently results in unnecessary biopsy and typically limits screening to patients with PSA values >4.0 ng/mL. The investigational ProCaM assay detects CpG island methylation within the promoter regions of three markers (GSTP1, RARß2, and APC) that are indicative of the presence of prostate cancer. The objective of this research study was to assess assay testing reproducibility and lot-to-lot variability. Methods: Assay reproducibility: 8 operators from 4 external clinical laboratories tested a panel comprised of a negative panel member (NM2C5 cells) for the internal control (ß-Actin), a high positive and low positive panel members (LNCaP cells) for all 3 markers and ß-Actin. Variability of assay results based on marker cycle threshold (Ct) values was determined. Lot variability: 3 unique lots of the ProCaM Test Kit were evaluated on DNA extracted from 169 post-DRE urine samples to show reproducibility across kit lots. Urine samples were collected from consenting subjects that were scheduled for prostate biopsy and tested by 3 operators at the same facility. Results: Assay reproducibility: The informative rate was 98% (766 valid results of 782 total tests). The overall intersite %CV and SD values for Cts were = 9.2% and 1.49%, respectively. The percent agreement with qualitative (positive/negative) outcome for High, Low, and Negative panel members was = 98% for GSTP1, RARß2, APC, and ß-Actin. Lot variability: Of the 169 biopsy results a histology positivity rate of 39% (66/169) was shown. Urine sample results for these samples showed that the areas under the curve for the 3 unique ProCaM Test Kit lots were equivalent (0.72, 0.74, 0.75, p > 0.263). Using the result categories of negative and positive with identical cutoffs for GSTP1, RARß2, APC for samples with >5 ssDNA copies 98% concordance was observed for all 3 lots evaluated. Conclusions: The investigational assay produces a qualitative result without the requirement for normalization or sample dilution and requires only one reaction per patient sample. Current results demonstrate that the assay has a high informative test rate and that results are reproducible across sites, operators runs, and kit lots.

[Table: see text]
Association of expression of bcl-2 with outcome in non-small cell lung cancer
Authors: Anagnostou V. , Lowery F. , Syrigos K. , Frangia K. , Zolota V. , Panagopoulos N. , Dougenis D. , Tanoue L. , Detterbeck F. , Homer R. , Rimm D.
Published: August 2020

e22039

Background: BCL-2 promotes cell survival by inhibiting adapters needed for the activation and cleavage of caspases thus blocking the proteolytic cascade that ultimately dismantles the cell. It is preferentially expressed in squamous cell carcinomas of the lung and has been investigated as a potential prognostic parameter in lung cancer patients with conflicting results. Here, we quantitatively assessed BCL-2 protein expression in two large and independent data sets to investigate the impact of BCL-2 on patient survival. Methods: AQUA, a fluorescent-based method for analysis of in situ protein expression, was used to measure BCL-2 protein levels and classify tumor by BCL-2 expression in a cohort of 180 lung cancer patients from Yale New Haven Hospital (training set). An independent cohort of 360 lung cancer patients from Sotiria General Hospital and Patras University Hospital in Greece was used to validate BCL-2 classification and evaluate outcome (validation set). Results: Tumors expressed BCL-2 in 57% and 53% of the cases in training and validation cohorts respectively and squamous cell carcinomas expressed higher levels of BCL-2 expression compared to adenocarcinomas (mean AQUA score 42 and 26 respectively, p=0.007); BCL-2 was not associated with other standard clinical or pathological characteristics. Survival analysis showed that patients with high BCL-2 expression had a longer median overall survival compared to the low expressers (20 vs 15 months, log rank p=0.016). Multivariate analysis revealed an independent lower risk of death for lung cancer patients with BCL-2 expressing tumors (HR=0.58, 95% CI 0.39–0.86, p=0.006). Conclusions: BCL-2 expression defines a subgroup of patients with a favorable outcome and may be useful for prognostic stratification of lung cancer patients as well as incorporation of BCL-2 into clinical decisions.

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Use of microRNAs to distinguish small from non-small lung cancer
Authors: Spector Y. , Meiri E. , Faerman A. , Ben David M. , Zepeniuk M. , Ezagouri M. , Cohen D. , Rosenwald S. , Perelman M. , Barshack I.
Published: August 2020

e22061

Background: Lung tumors are divided to two main classes: non-small cell lung cancer (NSCLC) that accounts for ∼80–85% of all lung primary tumors, and tumors from neuroendocrine origin - mainly small cell lung carcinoma and lung carcinoid. The classification of lung tumors can present a diagnostic challenge. New markers for different subtypes may aid in diagnosing difficult cases, can improve the accuracy of classification and could be important for selecting proper treatment. Here we studied the utility of microRNA as biomarkers for this differential diagnosis. MicroRNAs, a family of short non-coding regulatory RNAs, are highly tissue-specific and are well preserved in routinely prepared formalin-fixed, paraffin-embedded (FFPE) specimens, making them promising candidates as biomarkers for tissue and tumor classification. Methods: We used proprietary protocols for extracting high-quality RNA from FFPE samples. We used microRNA microarrays to profile more than a hundred samples from different histological subtypes of lung cancer including small cell, lung carcinoid and various types of NSCLC. Differential microRNA expression was verified using a microRNA qRT- PCR platform. Results: We found that several microRNAs are significantly differentially expressed between different subtypes of lung cancers. Specifically, using combinations of few microRNAs, we were able to accurately differentiate between neuroendocrine and NSCLC. Small cell and carcinoid tumors can be further distinguished using the signals of additional microRNAs, with very high sensitivity and specificity. Conclusions: Our results underscore the potential of microRNA expression for classification of tumor subtypes. We found that combinations of small numbers of microRNAs can successfully aid in the differential diagnosis of lung tumors, and provide a basis for the development of simple and reliable assays for clinical oncology.

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Development of ultra-sensitive single molecule immunoassays for the intracellular kinases Akt1/pAkt1 and Erk2/pErk2 for small tissue studies
Authors: Todd J. , Held D. , Klein B. K. , Macy E. , Agee S. J. , Lu Q. A. , Puskas R.
Published: August 2020

e22068

Background: The Ras/Raf/MEK/ERK and PI3K/PTEN/AKT kinase cascades are critical pathways in malignant transformation, tumor progression and multi-drug resistance (MDR) in human cancers. Expression and genetic studies have contributed much to our understanding of this role, but further clinical investigations at the post-transcriptional level are necessary. Towards this end, Singulex has developed single molecule immunoassays for total Akt1, pAkt1, total Erk2 and pErk2 utilizing the Erenna Immunoassay System for use in small tissue studies and clinical trials. Methods: Analytical sensitivity, inter- assay precision, linearity and specificity were determined and compared to corresponding ELISA assays. Novel assays were tested on lysates from a variety of commonly utilized cell lines in cancer studies: MCF-7, PBMC, Jurkat, HeLa, and NIH3T3 cells. Results: All four novel assays were significantly more sensitive than the ELISA assay method based upon sample endpoint dilutions. Acceptable precision (<20%CV) was demonstrated for the Akt1/pAkt1 assays in PBMC lysates and for the Erk2/pErk2 assays in MCF-7, HeLa and NIH3T3 lysates. The LLoQ of the Akt1/pAkt1 assay in Jurkat lysates was sufficient to quantify Akt1/pAkt1 from as few as 10–20 cells. Based upon a typical 1mm needle biopsy, this is sufficient to quantify Akt1/pAkt1 from less than 1/1000th of that biopsy. Conclusions: We show that the Singulex Akt1, pAkt1, Erk2 and pErk2 assays are over 50 fold more sensitive than currently available ELISA assays, and specifically quantify analyte in a variety of cell lysates. We have further demonstrated that the Akt1/pAkt1 assay can quantify from as little as 1/1000th of a typical needle biopsy. Thus, these novel assays greatly improve upon pre- existing technology, providing a new tool for clinical trials investigating Ras/Raf/MEK/ERK and PI3K/PTEN/AKT pathways previously limited by ELISA sensitivity.

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Evaluation of serum profiles changes after neoadjuvant chemotherapy for breast cancer using MALDI-TOF/MS procedure
Authors: Hawke D. , Mazouni C. , André F. , Baggerly K. , Baggerly K. , Tsavachidis S. , Buzdar A. U. , Martin P. , Kobayashi R. , Pusztai L.
Published: August 2020

e22072

Evaluation of serum profiles changes after neoadjuvant chemotherapy for breast cancer using MALDI-TOF / MS procedure. Background: Response to primary chemotherapy (CT) for breast cancer is heterogeneous among patients and a more tailored treatment would be beneficial in term of reducing exposure to an unnecessary toxicity and optimization of response rates. Mass spectrometry analysis of serum might be helpful in detecting specific changes in response to primary CT. Methods: An applied Biosystems 4700 Proteomics Analyzer matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometer was used. A breast cancer cohort of 78 sera samples from 39 HER2 positive patients consisting of matched pretreatment and (6 months) posttreatment samples was used. Blood samples were collected serially before each treatment cycle every 3 weeks of neoadjuvant CT. Samples were divided into those who achieved pathological complete response (pCR, n= 20) and those who had residual disease (RD, n=19). Low-mass differentially expressed peptides were identified using MALDI-TOF/TOF. Results: This procedure yielded a total of 2329 and 3152 peaks respectively, for the responders and non-responders. Biological variation analysis revealed a total of 32 peaks for responders and 643 peaks for non-responders to be differentially regulated with a false discovery rate less than 20%. A total of 8 differentially expressed proteins were identified from their peptides after digestion and LC-MALDI-TOF/TOF. Four in tumors with pCR (AFM, C3, hemopexin, SAP) and four proteins in the RD group were identified (AP1, hemopexin, Complement B, amyloid P component) Conclusions: Our study suggests that MALDI mass spectrometry may be used to predict the tumor response to neoadjuvant chemotherapy. Proteomic analysis may be useful in developing tailored chemotherapy for breast cancer.

No significant financial relationships to disclose.
Correlation of WT1 expression with tumor characteristics and clinical outcomes in mucinous breast carcinoma
Authors: Liang H. , Ye F. , Lembersky B. C. , Bhargava R.
Published: August 2020

e22075

Background: Wilms' tumor 1(WT1) protein expression has been evaluated by immunohistochemistry (IHC) in various histologies of series of invasive breast carcinomas. It is overexpressed predominantly in the mucinous subtype, in both pure and mixed mucinous carcinoma. The objective of this study was to correlate WT1 expression with the clinical and biological features of mucinous breast carcinoma as well as with the clinical outcomes and management. Methods: A total of 83 cases of pure and mixed mucinous breast carcinoma diagnosed between January 1999 and May 2006 were selected from the Magee-Womens Hospital, University of Pittsburgh Medical Center pathology registry. WT1 expression was examined by IHC using tissue microarrays (TMA). Only nuclear staining for WT1 was counted. Patient age, tumor features, management, and clinical outcomes were recorded, and survival was documented at the last patient followup. Results: Median follow-up was 59m (13–158m). The vast majority of patients had strong expression of hormone receptors. Only 3 patients had HER2-positive tumors. Disease free survival (DFS) was 92%. Overall survival (OS) was 88%. Sixty four cases had sufficient tissue on TMA for WT1 expression. Among them, WT1 expression was identified in 21/33 (64%) of pure mucinous carcinomas and 9/31 (29%) of mixed type tumors. Patients in both WT1-positive and WT1-negative groups had similar rates of breast conserving surgery, adjuvant radiotherapy and endocrine therapies. However, patients in the WT1-positive group were older (median age 74 vs 55 years, p=0.02) and had smaller tumor (median tumor size 1.2 vs 1.75 cm, p=0.03) as compared to those in WT1-negative group. More patients with WT1-negative tumors received adjuvant chemotherapy (40.6% vs 13.8%, p=0.02). Although all the recurrences were observed in patients with WT1-negative tumors, there was no statistical association of DFS or OS with WT1 expression. Conclusions: Overall, both pure and mixed mucinous breast carcinomas have excellent prognosis. This retrospective study demonstrated that WT1 expression may be associated with favorable outcome/pathologic features for patients with mucinous breast carcinoma. Prospective clinical studies will help to further elucidate these findings.

No significant financial relationships to disclose.