bluuebunny/crossref_metadata_2025 huggingface dataset:
Prevention of peristomal skin excoriation
Authors: Phadke M. V.
Published: August 2020

e20746

Background: Purpose of this paper is to prevent the peristomal skin excoriations. So far there has not been any permanent solution for this complication. Approximately 45% of patients with stoma suffer from this problem and are told to learn to live with it. It is important to understand the pathogenesis of any complication before a solution can be found. The excoriation results from digestion of the epithelium by enzymes in the mucus flowing out of the stoma. The liquid contents of the feces, because of their physical property, run along the mesenteric border of the intestine which is shorter than the anti-mesenteric border. This is complicated by bacterial contamination present in feces. It is essential to have intact mucosa all around the stoma. Skin is less resistant to bacterial invasion when compared with mucosa. When peristalsis starts, it tends to pull the mesenteric corner of the stoma inside the peritoneal cavity. This will continue till the mesenteric corner comes to the skin level. At this point there is no mucosa to protect and the liquid intestinal contents, instead of dropping in the bag leak over the skin and start digesting the epithelium. To change the mechanics of peristalsis, it is necessary to separate the mucosal (inner) tube from the serous (outer) tube. Once this is achieved, the peristalsis affects only the serous tube and has no effect on the mucus tube which forms the lumen of the stoma. This results in a 360 degree mucosal protection of the stoma. Methods: Delayed-Primary Self-Maturation a technique of constructing stoma described by Phadke was utilized in constructing all ileostomies and colostomies. Primary maturation of stoma is avoided. The stoma is left obstructed with staples. It is opened with electro-cautery only after good peristalsis has returned. The opening is at the anti-mesenteric corner. Mucosal cuff protrudes and is pushed by peristalsis. It everts and debrides the eschar of serositis, exposing the granulation tissue. The cuff rolls out and auto-grafts over the granulation tissue. The margins of the cuff come in contact with dermis of the opening in skin and complete the maturation process. Results: 63 Colostomies and 17 Ileostomies were constructed using this technique. All complications were prevented. Conclusions: DPSM technique will prevent peristomal skin excoriations and all other complications.

No significant financial relationships to disclose.
Efficacy and safety of administration of gemcitabine and docetaxel with radiation therapy in patients with high-grade soft tissue sarcoma
Authors: Raval G. G. , Govindarajan R.
Published: August 2020

e21503

Background: Neo-adjuvant therapy of soft tissue sarcoma is not standardized. Anthracyclines (A) improve disease free and overall survival in high grade extremity soft tissue sarcomas measuring more than 5 cm. A and ifosfamide (I) therapy require inpatient administration and is associated with significant toxicities when administered in combination with RT. G in combination with D has been shown to prolong progression free and overall survival in patients with advanced or metastatic STS after progression on A and I, and in patients unable to receive the combination. The safety of administration of G with radiation is not known. We evaluated the safety of neo-adjuvant G and D in combination with RT in patients with STS. Methods: Retrospective analysis of subjects with high grade STS treated with RT and neo-adjuvant chemotherapy with G and D was conducted. G 600mg/m2 was administered on days 1 and 8 along with D 75 mg/m2 on day 8 for a total of 3 cycles. 2200 cGY RT was administered in 11 daily fractions between cycles 1 and 2 and between cycles 2 and 3. 72 hour gap was given between RT and chemotherapy. Chemotherapy was administered on outpatient basis. Efficacy was evaluated by the extent of tumor necrosis at the time of surgery. Safety was evaluated by the presence of complications following chemotherapy. Results: GD + R was administered to 12 patients in the neo-adjuvant setting. 11 of them were evaluable for response. 10 of the 11 patients had evidence of tumor necrosis at the time of surgery. 1 patient had poor response. 3 of 11 patients had neutropenia complicating neo-adjuvant chemotherapy. 4 patients required hospital admission post chemotherapy. 3 for neutopenic fever, and 1 for fever, chills and pneumonia without neutropenia. Conclusions: Gemcitabine and taxotere combination with radiation therapy can be administered safely on outpatient basis with minimal toxicity. Further safety and efficacy evaluation of this therapy will need to be confirmed in a prospective phase II study.

No significant financial relationships to disclose.
Angiogenic biomaker study in osteosarcoma
Authors: Pavoni-Ferreira P. C. , Petrilli A. S. , Alves M. T. , Jesus-Garcia Filho R. , Toledo S. R.
Published: August 2020

e21507

Background: This study represents a prospective assessment of angiogenesis genes mRNA expression in tumors and blood from patients treated with pre- and post-operative Brazilian osteosarcoma protocol (GCBTO 2006) that introduce metronomic chemotherapy (anti-angiogenic) in order to try to increase survival of osteosarcoma patients. Methods: Tumor samples from 27 patients were analyzed before and after chemotherapy to determine VEGFA, VEGFR1, VEGFR2, PDGFC, SDF1 and TSP1 genes expression profile by Quantitative Real Time PCR. Also, blood samples of these patients were investigated pre- and post-chemotherapy and at the end of high-dose chemotherapy trying to evaluate potential for proangiogenic factors and antiangiogenic factor (TSP1) which could be used to monitor treatment activity. Results: Of all six genes studied pre- and post- chemotherapy, in tumor samples, only SDF1 and VEGFR2 were underexpressed. SDF1 gene has the lowest expression at all. In tumor samples, TSP1 and VEGFA expression was higher than SDF1, VEGFR2 and PDGFC expression in biopsies and surgeries (P=0.001). VEGFR1 expression was higher than VEGFR2 expression (P=0.001). PDGFC and VEGFR1 overexpression were associated with necrosis grade I and II (Huvos score) (P=0.005). VEGFA and TSP1 were overexpressed in 96% and 92% of surgery samples, respectively. In blood samples from biopsy, surgery and end of treatment there were no statistically significant changes in the marker genes expression. Conclusions: The study suggests an association between PDGFC and VEGFR1 overexpression and lower grade necrosis. TSP1 and VEGFA were the most expressed genes in all tumor samples but TSP1 was lower than VEGFA in biopsies and VEGFA was lower than TSP1 in surgery (P=0.001). Although VEGFR2 is the primary receptor of VEGF, VEGFR1 was the most expressed VEGF receptor.

No significant financial relationships to disclose.
GIST-RISK study: Risk of recurrence in primary resectable GIST
Authors: Lopez-Pousa A. , Artigas Raventos V. , Lucena de la Poza J. , Díaz de Liaño Arguelles Á. , Fernández Hernández J. , Bernal Jaulín J. , Talavera Eguizabal P. , Martí Obiol R. , Delgado Rivilla S. , Martínez Molina E.
Published: August 2020

e21514

Background: Several risk classification scales have been proposed to estimate the risk of recurrence after surgery for primary GIST, identifying different factors: location, tumor biology, the patients (pts), and type of surgery. Methods: Between June 2007 and December 2008 we performed a retrospective study on primary GIST pts, to analyze potential prognostic factors for recurrence, according to the risk classifications proposed by Fletcher and Miettinen, and the impact of clinical and treatment variables. Results: As of October 2008 a total of 79 pts were enrolled. Pts characteristics: male 59.5%; median age 69 years (27–90). Symptoms: abdominal pain 33%; hematemesis 32%; abdominal mass 11%; anemia 10%; non-symptoms 20.3%. Two pts received preoperative imatinib. Laparotomy was performed in 69.7%, laparoscopic resection in 27.8% and both in 2.5% of pts (2 pts intraoperative tumor rupture). A R0 resection was performed in 74 pts (93.7%), R1 in 3 pts (3.8%) and R2 resection in 2 (2.5%) pts. Tumor location: 54 gastric, 25 non gastric. Median size 5.4 cm (range 0.5–35). Tumor size: <5cm 46%, 5–10cm 39% and >10cm 15% of pts. Mitotic index (per 50 HPF): ≤5 mitosis 69%, 6–10 mitosis 15% and >10 mitosis 16%. Histology: spindle-shaped 63%, epithelioid 6%, mixed cells 11%, not available 19%. Inmunohistochemistry: CD117 + 95%. Mutational analysis was obtained in 5 pts (four exon 11; one exon 18 mutation). Adjuvant imatinib was administered to 13 pts (16.5%). According to Fletcher: high-risk 31,1%, intermediate-risk 29,7%, low-risk 28,4%, very low-risk 10.8% of pts. According to Miettinen: high-risk 19.4%, moderate-risk 9.7%, low-risk 27.9%, very low-risk 20.8%, none risk 11.1%, insufficient data 11.1% of pts. Conclusions: Preliminary results shows a different risk of recurrence according to NIH consensus or Miettinen scales, with more than 60% versus 30% of pts with intermediate-high risk of recurrence. This study is still ongoing, additional data and follow-up will be provided.

No significant financial relationships to disclose.
Hypofractionated radiation therapy for palliation of sarcoma metastases
Authors: Merimsky O. , Soyfer V. , Corn B.
Published: August 2020

e21517

Background: RT is our preferred modality for local palliation of metastatic STS, regardless of systemic chemotherapy. While a protracted course of RT may be given as postoperative adjuvant treatment, a short and intense course of RT is usually needed for rapid palliation and local control of metastatic disease. Sarcomas are usually considered, at best, as moderately radio responsive tumors. RT doses within the range of 60 - 70 Gy are usually needed to be delivered in order to eradicate microscopic disease, while 50 Gy doses are needed for other malignancies such as breast or rectal cancer. Methods: Seventeen patients, 8 women and 9 men, at a median age of 61 years (range 53–95 years) had symptomatic metastatic sarcoma, and required rapid palliation. In total there were 20 sites of involvement by metastatic disease: trunk (chest wall, groin, axilla)- 13 cases, limb- 7 cases. The symptoms related to the metastases were either pain or discomfort in all the patients. In 15 cases the RT was the only modality for local palliation and in 5 cases RT was given following metastasectomy with close or involved margins. All the patients were treated by a short and intensive course of administration: 39 Gy were given in 13 fractions of 3 Gy/day, 5 times a week. Results: Median follow-up period was 25 weeks. The treatment was well tolerated. Acute side effects included first-degree burn (skin redness) and mild superficial pain in all the cases. No wound complications were noted among those undergoing surgery. Late side effects included skin pigmentation and induration of irradiated soft tissues. Durable pain control was achieved in 12 out 15 cases treated for gross metastases. Tumor progression was seen in the 3 other cases within a period of 2 to 9 months. Among 5 lesions which were irradiated as an adjunctive treatment following resection, no local recurrence was observed. Conclusions: The results of this series, although limited in size, point to the safety and feasibility of hypofractionated RT for palliation of musculoskeletal metastases from sarcoma.

No significant financial relationships to disclose.
Chordoma: A rare bone tumor: A single institutional experience on a case series of 23 patients
Authors: Ferraresi V. , Nuzzo C. , Zoccali C. , Graziano V. , Graziano V. , Salducca N. , Zeuli M. , Giannarelli D. , Pellegrini D. , Cognetti F.
Published: August 2020

e21519

Background: Chordoma is a rare primary bone tumor arising from primitive notochord remnants of axial skeleton. Adequate surgery still remains the mainstay of its therapeutic treatment. Methods: Our review has been focused on the clinical characteristics, type of treatment, and prognosis of 23 consecutive cases of chordomas observed from 2004 until today and followed by the same team of specialists (orthopedic surgeons and medical oncologists). Results: From June 2004 to December 2008, 23 pts with various stage of disease went to our observation. Male/female: 18/5. Median age at diagnosis: 60 yrs (range: 40 -74). Sites of origin: sacrum (10 pts), spine (12 pts), and skull base (1 pt). Ten pts (44%) had primary disease, 9 pts (39%) had recurrent disease, and 4 pts (17%) had developed metastatic lesions. Surgery was part of the treatment in 21 out of 23 pts. Surgical margins were rated as wide/marginal in 17 pts and intralesional in 4 pts. Adjuvant proton radiotherapy has been delivered to 2 pts. Conventional radiotherapy was performed in 4 pts. The rate of local relapse after primary surgical excision was 76%. The median time to recurrence was 14 months (range: 2–113).Sixteen pts with inoperable/unresectable or metastatic PDGFR β expressing disease were treated with imatinib mesylate (IM) in the context of a clinical trial and a subsequent expanded access program (Stacchiotti S., proc ASCO 2007. abstr. 10003). One pts received weekly cisplatin in combination with IM after progression during IM therapy. The 5-year survival rate of the entire series was 76 %. Two out of 17 pts (12%) surgically treated with adequate margins are still alive and without evidence of disease; two other pts died without evidence of disease. Conclusions: Despite the progress of current surgical techniques and some encouraging results with the use of targeted therapy, chordoma results, generally, in a long-lasting disabling and life-affecting disease. Nevertheless, specific experience of the multidisciplinar team is an important prerequisite to improve patients’ quality of life and, hopefully, outcome.

No significant financial relationships to disclose.
Osteosarcoma in pediatric and young adult patients: The Singapore experience
Authors: Aung L. , Chan Y. H. , Tan A. M. , Wong P. S. , Pho R. W.
Published: August 2020

e21522

Background: There is limited information on treatment and survival regarding these tumors in the South East Asia region. We thus attempted to investigate the incidence, the treatment modalities used and the outcome of OS patients treated at the two major pediatric oncology hospitals in Singapore. Methods: A comprehensive list of patients with OS treated at the Department of Pediatrics National University Hospital and KK Women's and Children's Hospital, Singapore between April 1980 and May 2006 was generated. During the study interval, patients received neoadjuvant chemotherapy followed by definitive surgery consisting of either limb-salvage or amputation followed by adjuvant chemotherapy. Chemotherapy included combination of cisplatin and doxorubicin as per the European Osteosarcoma InterGroup (EOI) and as per the Memorial Sloan-Kettering Cancer Center's (MSKCC) T12 protocols. Treatment of subsequent relapses consisted of various combinations of methotrexate, ifosfamide, etoposide, other, and/or surgery. Results: Of the total 49 patients with OS, 14 (29%) patients presented with metastatic OS (lungs, n=13; skip lesion, n=1). The median age of diagnosis of OS was 12.4 (range, 6.6 - 17.9 years). Mean survival after diagnosis of OS was 13.4 years (95% CI, 9.1 - 17.8 years). The two and five-year overall survival were 56.4 % ± 86.9 % (se) and 37.5 % ± 73.9 % (se) respectively for the entire cohort. The two and five-year overall survival were 77.9 % ± 100 % (se) for patients who were treated as per the MSKCC T12 protocols. The two and five-year overall survival were 51.8 % ± 86.2 % (se) and 33.8 % ± 72.6 % respectively for patients who were treated as per the EOI regimen. Conclusions: Survival from OS in Singapore appears to be improving since the multi-drug combination chemotherapy was introduced in 2002. The rarity and complexity of OS makes it crucial for patients diagnosed with this tumor to seek a centralized multi-disciplinary Musculoskeletal Oncology team.

No significant financial relationships to disclose.
Prognostic factors in synovial sarcoma
Authors: Castillo Fernandez O. O. , Pacheco M. , Lim M. , Singh C.
Published: August 2020

e21525

Background: Synovial sarcoma (SS) is a relatively rare type of soft tissue sarcoma. Several prognostic factors have been described that modulate patients’ outcome. The aim of this study was to identify prognostic factors in patients with SS. Methods: A retrospective analysis was performed on clinical and histopathological data of patients treated at National Oncology Institute (Panama) between 1995 - 2007. Demographic, clinical, pathological, and therapeutic variables were reviewed. Overall survival and recurrence-free survival were analyzed using Kaplan-Meier and log-rank tests. Results: Twenty seven patients were analyzed, 56% male and 44% female, median age was 28. Tumour site; extremities based 85%, truncal 15%. Median tumour size was 8cm. Presentation; non-metastatic 84%, metastatic 16%. Histological type; monophasic 78%, biphasic 22%. Mitosis/10HPF; twenty and more 33%, less than twenty 67%. Necrosis; absent 63%, less than half 30%, and more than half 7%. Histological grade (FNCLLC); grade 2 48%, grade 3 52%. Surgery; limb sparing surgery 52%, amputation 48%. Treatment: adjuvant treatment 52% (3 chemotherapy only, 7 radiation therapy, 4 both). 90% were doxorubicin based regimens. The median follow up was 25 months. 63% of tumours recurred and 33% of patients died. Univariate analysis revealed a significant positive relationship between histological grade and overall survival (p<0.05). In patients with localized disease, time to recurrence decreased with histological grade (p=0.05 ns). Conclusions: High histological grade is an adverse prognostic factor for overall survival in patients with SS.

No significant financial relationships to disclose.
Primary breast sarcoma: Clinical and retrospective analysis of cases from Jerez General Hospital, Spain
Authors: Moreno-Vega A. , Chavarría N. , Rubio J. , Villandiego I. , Estepa R. , Gordon M. , Salvador J. , Jimenez E.
Published: August 2020

e21526

Background: Primary sarcomas of the breast (PBS) are a heterogeneous group of tumors from stromal breast, infrequent (0.1% of BC). Diagnosis and treatment is unclear. We analyzed all cases of PBS from BC diagnosed at our institution from 2004 to 2008. Methods: Histopathological specimens of PBS were reviewed (excluded phylloides, metaplastic and sarcomatoids carcinomas). A single-independent pathologist revised biopsies to confirm histology features (H&E) and inmunohistochemistry (IHC) setting: p53, ki67, vimentine, oestrogen and cerbB2 receptors, CD31/34, cito-keratins (CK) and histology grade. We revised image methods (mammography, ultrasound or IRM/TC) and surgery make. We analysed diseases outcomes (disease free survival, DFS) by histology high risk factors (tumor size, histology, and proliferation index). Heterogeneous chemotherapy/radiotherapy schedules was evaluated . Statistical analysis with G-Stat 2.0 was performed. Results: Seven cases of PBS (1 male/6 female) were reviewed, from 790 BC diagnosed (0.8%): 2 angiosarcomas (AS), 1 malignant fibrous histiocytoma, 2 undifferentiated, one osteoclastic and other spindle-cell sarcoma. Mean age was 63 years, 3 image-techniques used in 71.4% patients (pts), lumpectomy was performed in 71.4%. IHC: vimentine + and CK- in all cases, CD31/CD34+ in AS. P53 and Ki67 were not significantly associated with DFS (p=0.33 and p=0.88 respectively), tumor size (mean 7.4 cm) neither (T-Student test, p=0.80). Adjuvant therapy was radiation 57.14% pts; and chemotherapy (doxorubicin 4/liposomal doxorubicin 2 pts) in recurrence. Three pts have died with distant metastases (DFS mean: 11.7 months; Kaplan-Meier method). Contralateral low grade AS in one woman, and lung adenocarcinoma in the man, had been diagnosed 2 years later. Conclusions: PBS are rare and difficult diagnosis tumors. There are few series published, without prospective studies to evaluate an adequate therapy, diagnosis and valuable prognostic factors. Our incidence was high, but the independent pathology analysis confirmed all histopathological diagnosis. This review included novel IHC and IRM images, considered necessary for diagnosis and personalized treatment.

No significant financial relationships to disclose.
Potential usefulness of OPG, a member of the tumor necrosis factor receptor superfamily, for the early diagnosis of bone metastasis in breast cancer patients
Authors: Ibrahim T. , Sacanna E. , Mercatali L. , Flamini E. , Tison C. , Ricci R. , Ricci M. , Serra P. , Scarpi E. , Amadori D.
Published: August 2020

e22000

Background: Breast cancer is the most frequent tumor in women. About 80% of patients with metastatic disease present bone involvement, in which the OPG/RANKL/RANK system would seem to play an important role. Our aim was to evaluate the potential usefulness of OPG bone marker for the early diagnosis of bone metastases. Methods: The study was carried out on 120 individuals: 30 healthy donors, with a median age of 40 years (21–76) and 90 breast cancer patients, with a median age of 57 years (30–86). Among patients, 49 were disease-free (median age 52 years) and 41 were at first diagnosis of bone metastases (median age 63 years). OPG transcript was determined in peripheral blood samples using quantitative PCR analysis. A receiver operating characteristic (ROC) curve was used to calculate the diagnostic accuracy of the marker. Results: OPG values were not correlated with age in any of the subgroups. The OPG median value was not statistically different in healthy donors (median=1.9; range 0.6–4.7) and disease-free patients (median=1.7; range 0.4–8.9), whereas it was threefold higher than that observed in relapsed patients (median=0.6; range 0.1–5.2; p<0.001), regardless of the number of metastatic sites. The area under the curve (AUC) in disease-free patients was 0.82 for OPG, with 71% sensitivity and 88% specificity, using a cut off ≤ 0.8. In a parallel analysis of 37 patients (14 disease-free and 23 with bone metastases) for whom CEA and CA15.3 information was available, specificity for each marker was 100%, whereas sensitivity was only 61% and 59%, respectively. When these markers were considered in combination with OPG, an increase in sensitivity, albeit not statistically significant, was observed for CEA (83%) and CA15.3 (82%). Conclusions: Our preliminary data show a potential role of the OPG bone turnover marker for the early diagnosis of bone metastases. Results now need to be confirmed in a larger case series.

No significant financial relationships to disclose.
Cytogenetic abnormalities in mesenchymal stem cells in chronic lymphocytic leukemia (CLL) patients and normal subjects
Authors: Ding W. , Knox T. R. , Smoley S. A. , Van Dyke D. L. , Kay N. E.
Published: August 2020

e22002

Background: Mesenchymal stem cells (MSC) residing in the marrow support hematopoiesis and protect cancer cells from undergoing cell death induced by chemotherapy. Recent reports have described clonal cytogenetic abnormalities in the MSC of acute myeloid leukemia and myelodysplastic syndrome patients. To determine if cytogenetic abnormalities are present in MSC from CLL patients, we analyzed karyotypes of MSC from 13 CLL patients and 5 normal subjects. Methods: Stromal cells from marrow core biopsies of 13 CLL patients and 5 normal control subjects were isolated, cultured and confirmed as MSC based on their immunophenotype and capacity to differentiate into three lineages. After 3–4 non-stimulated cell culture passages, the karyotype was analyzed in 5–40 metaphase cells from each subject Abnormalities were considered clonal using the accepted convention of the same chromosomal gain or rearrangement in 2 or more cells or loss in at least 3 cells. For each CLL patient, interphase FISH analysis to detect the common CLL-associated abnormalities was performed on freshly isolated peripheral blood mononuclear cells (PBMC). Results: Clonal cytogenetic abnormalities of the cultured MSC were observed in 6 of 13 CLL patients (46%) and 3 of 5 control subjects (60%). The 6 CLL MSC had different clonal abnormalities than the PBMC CLL FISH studies. One CLL MSC exhibited trisomy 5, one exhibited trisomy 8, and one had monosomy X clone. One MSC had a balanced t(1;16). One MSC had a balanced t(3;12) and a del(3p), and one had a complex karyotype with 4 unbalanced rearrangements. The clonal aberrations observed in the 3 controls included a del(3p) in two cases, and in one case an inv(5) and a del(10q). There was no correlation of the chromosomal abnormalities in CLL MSC with clinical stage. Conclusions: Marrow MSC derived from CLL patients and normal subjects do show an array of cytogenetic abnormalities including clonal chromosomal abnormalities. However the genetic abnormalities found in both CLL and normal MSC could represent acquired genomic instability associated with advanced age, rather than oncogenesis associated with CLL.

[Table: see text]
The anatomic localization of circulating tumor cells and the immediate impact of surgery and radiofrequency ablation
Authors: Jiao L. , Apostolopoulos C. , Jacob J. , Johnson N. , Tsim N. , Habib N. , Coombes R. , Stebbing J.
Published: August 2020

e22004

Background: There are few data on the impact of immediate and differing surgical interventions on circulating tumor cells (CTCs), nor their compartmentalization or localization in different anatomic vascular sites. Methods: CTCs from consecutive patients with colorectal liver metastases were quantitated prior to and immediately after open surgery, laparoscopic resection, open radiofrequency ablation (RFA) or percutaneous RFA. For individuals undergoing open surgery, either hepatic resections or open RFA, CTCs were examined in both systemic and portal circulation by measuring CTCs in samples derived from the peripheral vein, an artery, the portal vein and hepatic vein. Results: A total of 29 consecutive patients with colorectal liver metastases were included with a median age of 55 (range 30 - 88 years). CTCs were localized to the hepatic portosystemic macrocirculation with significantly greater numbers than in the systemic vasculature. Surgical procedures led to a statistically significant fall in CTCs at all sites measured (p<0.05). Conversely, RFA, either open or percutaneous was associated with an increase in CTCs. Conclusions: Surgical resection of metastases but not RFA decreases CTC levels. In patients with colorectal liver metastases, CTCs are localized to the hepatic (and probably pulmonary) macrocirculations. This may explain why metastases in sites other than the liver and lungs, are infrequently observed in cancer.

No significant financial relationships to disclose.
Scotin expression and survival in breast cancer
Authors: Bourdon J. , Diot A. , Vojtesek B. , Bray S. , Jordan L. , Lane D. P. , Thompson A.
Published: August 2020

e22006

Background: The Scotin gene (3p21.3) is directly trans-activated by p53 in response to ionising radiation and produces a 25kD trans-membrane protein located in the endoplasmic reticulum and nuclear membrane which induces a caspase- dependent apoptosis. This study examines the expression of scotin and scotin isoforms in breast cancer in relation to clinical and pathological parameters. Methods: Scotin was examined in 145 breast cancers by DNA sequencing, nested PCR for mRNA expression and using novel antibodies to scotin (pantropic), the C-terminal or N-terminal domain, and the scotin-5 isoform with western blotting and immunohistochemistry on a tissue microarray of breast cancers. Results: Scotin was mutated in 10% of cancers and mRNA expression lost in a further 10%. Moreover, Scotin-5 was expressed in 40% of cancers, while it was not detectable in normal breast epithelium. We established that Scotin-5 is anti-apoptotic and inhibits scotin-mediated apoptosis. Cytoplasmic staining of scotin on TMAs demonstrated that loss of scotin expression was associated with earlier relapse (Mantel-Cox log rank p<0.01) and poor survival (Mantel-Cox log rank p<0.002). Furthermore, loss of Scotin expression identified 50% of the WTp53 patients or 50% of the ER+ patients who also had a poor survival. Conclusions: Scotin may be used as an independent biomarker in breast cancer. Loss of Scotin function may account for a significant proportion of p53 malfunction even in the presence of wild type p53 in breast cancer and adds a further level of complexity to p53 directed therapy in breast cancer.

No significant financial relationships to disclose.
Mammary adipose tissue and cancer cell growth: The role of adipose tissue in the tumor microenvironment
Authors: Carroll P. A. , Healy L. , Lysaght J. , Griffin M. , Dunne B. , Boyle Mr. T. , Reynolds J. V. , Kennedy M. J. , Pidgeon G. , Connolly E. M.
Published: August 2020

e22009

Background: Worldwide, the prevalence of obesity is rapidly increasing, correlating with a direct increase in cancer rates1. Adipose tissue is considered an important endocrine organ producing several important hormones and cytokines including leptin and adiponectin. Mechanisms for the role of obesity in cancer states includes the excess or unregulated secretion of adipocytokines from adipose tissue, and potentially the metabolic syndrome (a cluster of co-morbidities linked to metabolic dysregulation). Mammary adipose tissue is proposed to play a vital role in the microenvironment of normal and tumour states within the breast2. Breast adipose tissue is a good candidate to investigate effects of obesity and metabolic disturbances on cancer states. Methods: Peritumoural (PT) adipose tissue adjacent to the tumour and distal adipose tissue (D) within the breast was sampled in 10 patients. The tissue was processed and cultured for 72hrs in serum free minimal cytokine media. A-MB-231 and MCF-7 breast cancer cell lines were then cultured with this adipocyte conditioned media (ACM) and cell proliferation response was then measured using BrDU assays. The adipocytokine profile at the mRNA and protein level was measured in ACM and adipose tissue for comparative differences using RT-PCR, ELISA and Cytokine Profiler technology. Results: ACM from both sites promoted tumour cell survival. There was however, a differential cell proliferation response noted between the peritumoural (PT) ACM and that from the distant (D) ACM. Pro-inflammatory mediators (VEGF, TNF-α, EGF) were also demonstrated a trend to be differentially expressed between the 2 sites. Conclusions: Proliferation of breast cancer cell lines occurs in response to ACM, with differential effects seen between peritumoural ACM and distant ACM. This may be mediated through increased pro-inflammatory or pro- mitogenic adipocytokine production in adipose tissue surrounding tumour. Further analysis will determine what role obesity and the metabolic syndrome plays in the results noted.

No significant financial relationships to disclose.
HIF expression and Max-SUV-18F-FDG-PET in non-small cell lung cancer (NSCLC) patients
Authors: Lopez R. , Gallardo E. , Ruibal A. , Leon L. , Sanchez-Salmon A. , Abdulkader I. , Gude F. , Curiel T. , Barandela J. , Gayoso L.
Published: August 2020

e22010

Background: Tumor hipoxia induces the up-regulation of several genes via the hipoxia-inducible transcription factors (HIF) 1 and 2. HIF-2 alpha (HIF-2 α) and HIF-1 alpha (HIF-1α) are associated with the prognosis of operable NSCLC patients. We studied the immunohistochemical expression of HIF-1α and HIF-2 α in patients with NSCLC and the possible correlation with the maximum standardised uptake value (max SUV) of 18F-FDG as well as other biological parameters. Methods: We used a Tissue Arrayer device (Beecher Instruments. WI) to construct a TMA block, according to conventional protocols for the study of immunohistochemical expression of HIF-1α, HIF-2 α, EGFR, bcl-2, MIB1, p16, p63 and cyclins A, B1, D1 and D3. Sections were scored as positive if >10% of cells stained positively. Staining patterns were correlated to clinical variables. Results: HIF- 1α expression was observed in 84/96 patients (34/39 adenocarcinomas and 50/57 squamous cell carcinomas), however it did not correlate with clinical stage (I-II: 41/45 vs III-IV: 44/51). HIF-1α correlated positively with HIF-2 α (p:0,001) and EGFR (p:<0,001) expressions. The maxSUV values of 18F-FDG-PET were higher (p:0,039) in HIF-1α -positive (17,1±8,6) than in negative tumors (11,8±4,4). HIF-2 α expression was observed in 60/103 cases (27/42 adenocarcinomas and 33/61 squamous cell carcinomas) and it did not correlate with clinical stage ((I-II: 28/45 vs III-IV: 32/57). HIF-2 α correlated positively with HIF-1α (p:0,001), MIB1 (p:0,045) and EGFR (p:0,091) expressions. After multivariate analysis, only the clinical stage (RR: 2,2) was a prognostic factor. Conclusions: 1) HIF-1α and HIF-2 α expressions are frequent in patients with NSCLCs and it did not correlate with clinical stage; 2) maxSUVs FDG-PET values were higher in HIF1alpha positive than in HIF-1α negative patients; 3) HIF-1α was correlated with EGFR expression, while HIF-2 α was correlated with MIB1 expression.

No significant financial relationships to disclose.
The incidence of low-level HER2 gene amplification and chromosome 17 polysomy in cases of newly diagnosed breast cancer and their association with clinical prognostic markers
Authors: Diamond J. R. , Amaria R. , O'Donnell C. I. , Meltesen L. , Swisshelm K. , Melancon D. , McGavran L. , Singh M. , Elias A. , Borges V. F.
Published: August 2020

e22012

Background: The purpose of this study is to describe the frequency of low level HER2 gene amplification and chromosome 17 polysomy, and its association with known clinical prognostic markers. Methods: We retrospectively reviewed all cases of newly diagnosed invasive breast cancer referred to the Colorado Cytogenetic laboratory for FISH testing from either the University of Colorado Breast Center or St. Mary's Hospital in Grand Junction, CO between October 2000 and May 2005, excluding patients treated with HER2-directed therapy. Cases were classified into positive for HER2 gene amplification (FISH ratio ≥ 2.2), negative for HER2 gene amplification (FISH ratio <1.8), or intermediate for HER2 gene amplification (FISH ratio of 1.8- 2.1 or presence of chromosome 17 polysomy). IRB approved chart review was performed . Results: Of the 177 patients referred for FISH testing during the study period, 27.7 % (49/177) had intermediate HER2 gene amplification as defined as a FISH ratio of 1.8–2.1 or chromosome 17 polysomy. There was a statistically significant increase in stage, lymph node involvement, and histologic grade at diagnosis in the intermediate HER2 group compared to negative controls ( Table ). Conclusions: The incidence of intermediate HER2 gene amplification is clinically significant and associated with higher histologic grade and stage at diagnosis compared to HER2 negative controls. This patient population should be investigated further for long term disease outcomes and impact of HER2 - based targeted therapy.

[Table: see text]

No significant financial relationships to disclose.
Investigation of HDAC inhibition targeting of BRCA1 expression as a mechanism to enhance platinum sensitivity in breast and ovarian cancer
Authors: Weberpals J. I. , O'Brien A. , Garbuio K. , Clark-Knowles K. , Dimitroulakos J.
Published: August 2020

e22017

Background: The improved outcome of Breast-Cancer 1 (BRCA1)-deficient breast and ovarian cancer may be linked to the impaired ability to repair double strand breaks caused by DNA-damaging chemotherapy (CTX), such as platinum compounds. Therapeutically relevant agents that target BRCA1 expression to sensitize tumors to platinum have not been identified. In this study, we explore the effect of histone deacetylase inhibition (HDACi) on platinum sensitivity and BRCA1 expression in a breast and ovarian cancer cell line model. Methods: The efficacy of HDACi to potentiate the cytotoxicity of platinum-based chemotherapeutics was evaluated in a range of breast and ovarian tumor cell lines using the MTT cell viability assay and confirmed by flow cytometry. BRCA1 mRNA and protein expression was determined by Q-PCR and Western blot, respectively. The effect on DNA damage was measured by immunofluorescence staining and flow cytometry for γH2A.X foci, a hallmark for the presence of DNA double strand breaks. Results: Baseline BRCA1 expression was variable in two ovarian (A2780s, cisplatin-sensitive and A2780cp, cisplatin-resistant) and four breast cancer cell lines (MCF7, T47D, BT549 and HCC1937) with minimal and absent protein expression in BT549 and HCC1937, respectively. The addition of the HDACi, M344 increased the sensitivity of cells to cisplatin and carboplatin treatment in those cell lines with significant BRCA1 levels. Expression of BRCA1 protein decreased in response to the addition of HDACi to platinum in all cell lines. BRCA1 mRNA levels decreased with the addition of HDACi to platinum in all breast cancer lines and in A2780cp. A2780s and MCF7 cells subjected to combination platinum and HDACi treatment demonstrated increased levels of DNA damage, as assessed by the presence of phosphorylated γH2A.X foci. Conclusions: This study supports a novel mechanism of HDAC inhibition to sensitize breast and ovarian cancer cells to platinum via inhibition of the DNA repair protein BRCA1. BRCA1 expression changes may represent a novel biomarker to assess the activity of this combinational therapeutic approach in clinical evaluations of breast and ovarian cancer patients.

No significant financial relationships to disclose.
Expression profile of hepatic drug transporters in patients with hepatocellular carcinoma
Authors: Higuchi K. , Terada T. , Ogasawara K. , Aoki M. , Fukudo M. , Katsura T. , Hatano E. , Ikai I. , Inui K.
Published: August 2020

e22026

Background: Hepatocellular carcinoma (HCC) does not respond sufficiently to chemotherapy. It has been demonstrated that several transporters can cause drug resistance in tumor cells. In human liver, various drug transporters are expressed and play pivotal roles in hepatic uptake and excretion of drugs. In HCC, the expression profile of hepatic drug transporters may be altered due to pathophysiological changes. However, limited data are available on their expression and regulation in HCC tissues. The aims of this study were to determine the expression levels of hepatic drug transporters in HCC patients and to identify transporter specifically regulated in HCC tissues. Methods: The mRNA expression levels of 17 drug transporters, including 9 solute carrier (SLC) transporters (OAT2, OAT7, OATP1B1, OATP1B3, OATP2B1, OCT1, OCTN2, MATE1 and PEPT1) and 8 ATP-binding cassette (ABC) transporters (R1, MRP1–6 and BCRP), were determined by real-time RT-PCR in cancerous and non-cancerous liver tissues from 57 patients with HCC. Protein expression level was evaluated by Western blot analysis. In addition, DNA methylation status in promoter region of transporter gene was analyzed using bisulfite sequencing. Results: The mRNA expression levels of 11 transporters (OAT7, OCTN2, MATE1, PEPT1, R1, MRP1–6) were significantly increased (1.45 to 4.74-fold, p<0.05) in HCC tissues compared with non-cancerous liver tissues, whereas only OCT1 mRNA was significantly decreased (0.61-fold, p=0.0004). Notably, MRP4 mRNA showed the highest increase, which was consistent with the marked augmentation in the protein level. The frequency of methylation in MRP4 promoter region was comparable between HCC and non-cancerous liver tissues, suggesting other mechanisms than DNA methylation for MRP4 up-regulation. Interestingly, the mRNA expression levels of MRP4 did not change with the differentiation of HCC. Conclusions: It was revealed that most of the hepatic drug transporters were elevated in HCC tissues. Furthermore, we first identified that MRP4 showed remarkable increase in protein as well as mRNA expression levels. These findings suggest that MRP4 is a novel candidate for diagnostic marker and a target molecule to reverse multidrug resistance in chemotherapy for HCC.

No significant financial relationships to disclose.
Prognostic significance of serum vascular endothelial growth factor per platelet count in gastric cancer patients
Authors: Seo H. , Sung H. , Choi I. , Oh S. , Seo J. , Shin S. , Kim Y. , Park K. , Kim J.
Published: August 2020

e22031

Background: Recent studies have shown that VEGF expression not in tissues but in serum sample is correlated with tumor vascularity, and high serum VEGF levels could predict poor prognosis in cancer patients. However there are limited data regarding the clinical and prognostic significance of serum VEGF levels per platelet count in advanced gastric cancer. In this study, we conducted a study to evaluate the prognostic implication of serum VEGF per platelet count in patients with advanced gastric cancer. Methods: 111 patients with histologically confirmed gastric cancer, 10 patients with early gastric cancer were included and control serum samples were acquired from 10 healthy volunteers. The levels of VEGF were measured using human VEGF quantitative enzyme-linked immunosorbent assay (ELISA). Survival curves were calculated using the Kaplan-Meier method and survival comparisons were made by the log rank test in metastatic gastric cancer. The Cox proportional hazards regression model was utilized for multivariate analyses after univariate analysis defined relevant prognostic variables. Results: A trend toward a significant positive correlation between serum VEGF and platelet counts was observed in patients of AGC (r = 0.477, P = 0.000) and there was a significant correlation between serum VEGF levels and differentiation of tumor (p = 0.014), stage (p = 0.036). The overall survival (log rank, p =0.0432) and the progression free survival (median 4.5 vs. 8.9 months; log rank, p =0.0116) were significantly shorter in patients with high VEGF per platelet count (≥1.626 pg/106). In the multivarivate analysis, performance status (P=0.025), the presence of peritoneal carcinomatosis (P=0.006), serum VEGF per platelet (P=0.005) were found to be significantly associated with poor progression free survival. Conclusions: This study demonstrated that serum VEGF per platelet count are correlated with poor overall survival and progression free survival in patients with advanced gastric cancer. Therefore measurement of serum VEGF per platelet might be useful markers for predicting disease progression and prognosis of advanced gastric cancer.

No significant financial relationships to disclose.
Incomplete surgical resection of DCIS results in activation of HER-2 in residual breast cancer cells
Authors: Singer C. F. , Hudelist G. , Fuchs E. , Köstler W. , Fink-Retter A. , Gschwantler-Kaulich D. , Gnant M. , Lamm W. , Lamm W. , Rudas M. , Kubista E.
Published: August 2020

e22035

Background: HER-2 amplification and consecutive overexpression is a predictor for poor prognosis in breast cancer patients. In addition, incomplete resection of HER2 overexpressing tumors leads to increased proliferation of residual breast cancer cells. While the local release of cytokines is thought to be responsible for the malignant behaviour of remaining tumor tissue, the exact mechanism is still unknown. Methods: We have analyzed EGFR, activated (p)EGFR, and activated (p)HER2 protein expression in HER2 overexpressing and in non-HER2 overexpressing tumors from patients who underwent breast surgery and consecutive reexcision for involved margins, and compared expression levels by IHC. Results: While overall HER2 protein expression in the initial and the reexcised sample were comparable, we observed an increase in pHER2 in DCIS in both, HER2 overexpressing (16/21 vs 24/24; p=0.018, Chi Square test) and non-HER2 overexpressing tumors (3/28 vs 5/12; p=0.025, Chi Square test). pHER2 was not increased in invasive tumors, regardless on whether the samples had been taken from a HER2 overexpressing (9/25 vs 6/17; p=0.261, Chi Square test), or a non-HER2 overexpressing tumor (1/27 vs 0/8; p=0.581, Chi Square test). EGFR expression was only detected in 1/47 HER2 overexpressing primary tumors and 2/48 non-HER2 overexpressing tumors, and was undetectable in reexcised specimen. Conclusions: Taken together, we have demonstrated an increase in HER2 receptor activation in incompletely resected preinvasive breast cancer. We hypothesize that receptor phosphorylation is caused by growth factor stimulation in response to intraoperative tissue damage, and perioperative inhibition of specific cytokines could become a promising therapeutic strategy.

No significant financial relationships to disclose.
Molecular profiling in patients (pts) with upper gastrointestinal (UGI) cancers correlated with clinical outcome
Authors: Virk N. , Yang D. , Lenz H. J. , El-Khoueiry A. B. , Danenberg K. D. , Iqbal S.
Published: August 2020

e22042

Background: Recent efforts have expanded our understanding of the molecular pathogenesis of UGI cancers and how oncogenes and tumor suppressor genes play a role in both carcinogenic and metastatic processes. These markers may serve as prognostic and/or predictive factors for recurrence following resection and resistance to radiotherapy and chemotherapy. Gene expression levels of thymidylate synthase (TS), thymidylate phosphorylase (TP), epidermal growth factor receptor (EGFR) and excision repair cross complementing (ERCC-1) have been shown to be associated with outcomes in lung, gastric and colon cancer. We evaluated TS, ERCC-1, EGFR and TP gene expression levels in patients with UGI cancers. Methods: This was a retrospective study of 80 pts with UGI cancers evaluated at USC who underwent molecular profiling. The primary objective was to determine a correlation between TS, TP, ERCC1 and EGFR and correlate with clinical outcome. Characteristics of these 80 pts: (16 females, 64 males ); median age 61 years (range 34–85); tumor types evaluated - 32 (40%) esophageal, 24(30%) gastric, 24(30%) GE junction; stages- 53 % IV, 21% III, 12% II, 2% I were evaluated for intratumoral gene expression of TS, TP, ERCC-1, & EGFR by real time quantitative PCR using Taqman technology from microdisected paraffin-embedded tumor sections. Results: High TS expression was associated with shorter OS (12.8 months vs. 23.7 months p=0.036). A significant correlation was found between TP & ERCC-1 (p=0.0078, r=0.37); TP & TS (p=0.0128, r=0.35); TP & EGFR (p=0.0065, r=0.39); TS & ERCC-1 (p=0.0004, r=0.39); ERCC-1 & EGFR (p=0.025, r=0.30). No statistically significant relationship was found between TS & EGFR (p=0.06,r=0.25). There was no correlation of ERCC-1, TP, & EGFR with OS that reached statistical significance. Conclusions: TS mRNA levels were shown to be associated with OS in UGI tumors, consistent with data reported in colon cancer. TS gene expression was significantly associated with expression levels of ERCC-1. In addition, ERCC1 was associated with EGFR. These data show for the first time that molecular pathways of cytotoxic agents are linked to the EGFR pathway suggesting that sensitivity to fluoropyrimidines, oxaliplatin, and EGFR inhibitors may be associated.

No significant financial relationships to disclose.