bluuebunny/crossref_metadata_2025 huggingface dataset:
Burden of chemotherapy-induced febrile neutropenia in paediatric oncology in developing countries: The Children's Hospital Lahore Pakistan experience.
Authors: Ahmad Alia
Published: September 2018

e22021

Background: The Oncology Department Of The Children’s Hospital Lahore is a 60 bedded unit providing free treatment to over 1000 new childhood cancer cases each year.The aim of this prospective study was to analyze the burden of chemotherapy induced FN and to assess the leading risk factors . Methods: Prospective review of 100 patients with FN admitted in July to August 2016 was done. Data regarding their age, sex, and clinical features, baseline CBC, course of therapy, hospital stay and understanding of caregivers regarding FN analyzed. Results: Total 100 patients with age ranging from < 1 to 15 years (67% < 5 yrs) were included.M: F Ratio was 1.7:1, 72% of cases had ALL and 28% with solid tumors. 28% had last chemotherapy received in 72 hours, 30% in last week and rest in more than a week time 36% had respiratory tract infections, 18% gastrointestinal infections, 20% mucositis, 10% no focus found and rest 16% had other manifestations Only 2 % presented in < one hour of start of symptoms, 27% < 24 Hours, 61% in < 5 Days and 10% > 5 days duration of symptoms. 45% had Hb < 8 gm%, 33% had platelets < 50000, and 54% had WBC < 1000 and 63% had ANC <100 (p-Value = 0.003- Hospital Stay). 29% presented with first episode while 51 % had 3 or more FN episodes.28% cases stayed 1 hour distance from CHL while 72% had to travel > 1-5 hrs to reach the primary treatment center. 66% received paracetamol at home, 17 had oral antibiotics while 17% had no treatment before reached PTC. Only19% caregivers had adequate awareness regarding FN, 72% had some understanding while 9% had no knowledge about FN (p-value = 0.000). 40% had social issues, 41% were unaware while 13% showed negligence in seeking treatment .46% stayed for 5 days and 48% for more than 5 days. 21% patients had increased doses of oral chemotherapy not adjusted timely according to CBC. Conclusions: Health educationof caregivers and health professionals is of utmost importance to prevent and manage FN effectively. Need of capacity building in pediatric oncology in low income countries along with sustainable social support is essential for provision of standard childhood cancer care and implementation of effective infection control measures.
The open, multicenters, double-arms, randomized, parallel controlled clinical trial of lobaplatin or gemcitabine combined with docetaxel as second-line therapy for advanced osteosarcoma-the effectiveness and safety analysis.
Authors: Sun Yuanjue , Yao Yang , Cai Zhengdong , Yu Xiuchun , Wu Sujia , Ye Zhaoming , Hua Yingqi , Zheng Shui'er , Chen Guojing , Hu Yongcheng , Zhou Yong , Wang Zhen
Published: September 2018

e22506

Background: to study the progression free survival (PFS), objective response rate (ORR), overall survival (OS) and safety of lobaplatin or gemcitabine combined with docetaxel as second-line therapy for advanced osteosarcoma. Methods: the NO. of clinical trial was HNCA001,a total of 15 cases were enrolled into experiment group adopted lobaplatin combined with docetaxel,and 14 into control group received gemcitabine combined with docetaxel.The trial concluded screening, treatment and follow-up periods.21 dyas as one treatment period,they got drugs at the first day,and were evaluated the effects per two periods, treated at most 6 periods. Results: The baseline characteristics of two groups were no ststistical differences(diastolic blood pressure excluded) (P > 0.05).The PFS rates of 2,4 and 6 periods in experiment group were 87.50%、87.50% and 70.00%,83.33%,27.78% and 0 in control group,middle PFS more than 12 months in experiment group and 3.20 months in control group;while there was no statistical difference of PFS rates(χ2= 2.42,P = 0.1194).The survival rates of 2,4 and 6 periods in experiment group were 100.00%,83.33% and 83.33%,83.33%,83.33% and 0 in control group,the middle OS more than 12 months in the two groups, there was no statistical difference of survival rates (χ2= 0.65,P = 0.4196).The ORR was 6.67% in experiment group and 0 in control group(P = 1.0000).There were no adverse events occurring in the two groups. Conclusions: There is negative results of the trial,most reason may be the samples are greatly less;the next step is samples expanded and follow-up extended for the priority of lobaplatin or gemcitabine combined with docetaxel as second-Line therapy for advanced osteosarcoma. Clinical trial information: NCT02099396.
Real-world outcomes of ponatinib in treatment of advanced gastrointestinal stromal tumors (GIST) after tyrosine kinase inhibitor (TKI) failure.
Authors: Heinrich Michael C. , McGarry Lisa , Kerstein David , Huang Hui
Published: September 2018

e22508

Background: Imatinib is standard front-line treatment for advanced KIT+ GIST. For patients whose tumors progress on imatinib, sunitinib is indicated, followed by regorafenib for those with disease progression on both imatinib and sunitinib. Patients refractory to 2 prior TKIs typically experience rapid progression (median PFS: 4.8 mos for regorafenib; 0.9 mos for placebo [Demetri Lancet 2013]), and with failure of all approved TKIs, unmet need remains high. The oral TKI ponatinib, approved for TKI-refractory CML and Ph+ ALL, has shown activity in heavily-pretreated (76% with ≥4 prior regimens) advanced KIT+ GIST patients in a Phase 2, single-arm study (Heinrich ASCO 2015) at starting dose of 45 mg/day: clinical benefit rate (CBR = CR + PR + SD for ≥16 wks) of 37% and median PFS of 4.3 mos were observed in 30 patients with KIT exon-11-mutant GIST and CBR of 14% and median PFS of 2.0 mos in 15 patients whose GIST lacked a KIT exon 11 mutation. Methods: To examine real-world outcomes, we retrospectively identified patients receiving ponatinib off-label for GIST (ICD-9 171.5x, 171.9x) between 01Jan2014 and 31Dec2016 from sole-source US specialty-pharmacy records. Using patient, physician and pharmacy dispensing data, patient characteristics, treatment setting and dosing were noted, and duration of treatment (DOT) was examined using Kaplan-Meier techniques. Results: We identified 26 probable GIST patients receiving ponatinib over this 2-year period: 58% male; median age 57.5 yrs. Where reported, all received prior TKIs, but data for prior lines of therapy were incomplete. Most (69%) were treated in an academic setting, and the most frequent starting dose was 45 mg (77%). DOT ranged from < 1 to > 19 mos, with an estimated median DOT of 3.8 mos. At 3 mos, 61% of patients were estimated to remain on ponatinib, and at 6 mos, 34%. Conclusions: Available real-world data show that patients treated with ponatinib for TKI-refractory GIST had a median DOT of almost 4 mos, with 1/3 remaining on therapy for > 6 mos. Using DOT as a surrogate for PFS, these results are consistent with Phase 2 study results, and suggest ponatinib may have activity in patients who have exhausted other treatment options.
Various fashions of laparoscopic resection for gastrointestinal stromal tumors in esophagogastric junction.
Authors: Xiong Wenjun , Wang Wei , Wan Jin
Published: September 2018

e22512

Background: Laparoscopic surgery for small (<5 cm) gastric gastrointestinal stromal tumors (GIST) is now widely performed. However, laparoscopic resection of GIST in esophagogastric junction is technically difficult. Herein, we introduce various fashion of laparoscopic resection for small GIST in esophagogastric junction. Methods: Retrospective review of 40 consecutive patients with small GIST in esophagogastric junction who underwent attempted laparoscopic surgery. GIST in esophagogastric junction was defined as that the distance of the upper border of GIST from esophagogastric line was less than 2 cm. Three fashions of laparoscopic resection were performed: fashion A, laparoscopic wedge resection using linear stapler; fashion B, laparoscopic complete resection by opening the stomach wall and the stomach wall incision was closed with suture; fashion C, laparoscopic proximal gastrectomy with pyloroplasty. The data of clinicopathologic characteristics, operative course and short-term outcomes were analyzed. Results: All procedures were finished successfully and no operative relatively complication was recorded. Tumor in 24/40 (60%) patients was located in greater curvature. 70.1% (17/24) of them received fashion A and others (7/24) underwent fashion B. Tumor in 16/40 (40%) patients was located in lesser curvature. 18.8% (3/16) of them underwent fashion C and others (13/16) underwent fashion B. The mean operative time was 97.4±21.3 min and the mean estimated blood loss was 20.5±10.4 ml. The mean first time of flatus was 39.2±10.0 hours and the time of fluid intake was 40.1±11.7 hours. The mean hospital stay was 4.2±1.3 days. The mean diameter of tumor was 2.7±1.0 cm. Conclusions: Laparoscopic surgery for small GIST in esophagogastric junction is safe and feasible. The selection of various laparoscopic resection fashions was according to the tumor location.
A phase II trial of Q3 week or weekly paclitaxel in combination with bevacizumab for metastatic or unresectable angiosarcoma.
Authors: Bui Nam , Kamat Nikhil , Ravi Vinod , Ganjoo Kristen N.
Published: September 2018

e22513

Background: Paclitaxel (P) and bevacizumab (B) are both agents that provide clinical benefit in advanced angiosarcoma. The objective of this study was to assess the efficacy and safety of P-B in two regimens: Q3 week or weekly P with B for patients with advanced AS. Methods: We conducted a multicenter phase II trial. Patients were to receive P 200mg/m2 IV with B 15mg/kg IV every 21 days (Regimen A) or P 90mg/m2 IV weekly D1, 8, 15 with B 15mg/kg IV D1 of a 28 day cycle (Regimen B) x6 cycles. Maintenance B followed at a dose of 15 mg/kg intravenously once every 21 days. The primary end point was 4 month non-progression rate (NPR, RECIST 1.1). Results: A total of 16 patients were entered in the study between June 2010 through September 2013. Sites of disease included soft tissue (n = 5), skin/scalp (n = 5), breast (n = 4), and viscera (n = 2). All four breast patients were radiation induced after treatment for breast adenocarcinoma. 11 were chemotherapy naïve while 5 had received prior systemic chemotherapy. Results are reported as both an aggregate as well as per regimen (Table 1). 4 month NPR was 62.5% with median overall survival 16 months and median progression free survival 5.06 months. Of the 16 patients, 11 made it to cycle 3 and thus were evaluable for response. There were 1 CR, 4 PR, 2 SD, and 6 PD. There were no grade 5 toxicities, ten grade 3 toxicities and four grade 4 toxicities. The most common grade 3 toxicity was neutropenia (3/10) with other notable toxicities including peripheral neuropathy, hypertension, and heart failure. Grade 4 toxicities included neutropenia, bowel perforation, atrial flutter, and acute coronary syndrome. The breakdown between the two regimens revealed comparable efficacy and safety with similar ORR, 4mNPR, mOS, mPFS, and G3/4 toxicities. Conclusions: P and B followed by maintenance B is active and has a tolerable safety profile. Q3 week and weekly paclitaxel appear similar in efficacy and safety. Clinical trial information: NCT01055028. [Table: see text]
Single-center experience with intimal sarcoma, an ultra-orphan, commonly fatal mesenchymal malignancy.
Authors: Van Dievel Julie , Sciot Raf , Delcroix Marion , Vandeweyer Raf , Debiec-Rychter Maria , Dewaele Barbara , Cornillie Jasmien , Van Cann Thomas , Meyns Bart , Schöffski Patrick
Published: September 2018

e22532

Background: Intimal sarcoma is a rare malignancy that, clinically and radiographically, often mimics pulmonary embolism. The intravascular tumor tends to disseminate rapidly and metastases can be present at first diagnosis. Methods: We reviewed all cases of intimal sarcoma which were diagnosed, treated and followed at the University Hospitals Leuven between April 2006 and April 2016. Results: We identified 13 patients with a median age of 51. In 6 patients initial findings were suggestive of thromboembolic disease. PDGFRA amplification was the most prevalent molecular finding, present in 11 patients. The MDM2 gene was amplified in 9 cases, the EGFR gene in three patients. The median overall survival was 13 months. Eleven patients underwent surgery. In 5 cases with inoperable and/or metastatic disease chemotherapy was given. Treatment with imatinib was initiated in 4 patients. Conclusions: Intimal sarcoma is an extremely rare and aggressive malignancy that has a very poor prognosis. Mimicking thromboembolic disease, diagnosis and treatment can be delayed. Surgery is the mainstay of treatment but is seldom curative. The disease is highly resistant to cytotoxic and targeted treatment. Given the fact that intimal sarcoma commonly expresses more than one molecular target, combination therapy might be an option, though toxicity may be a limitation.
Complex approach in the treatment of retroperitoneal fibrosarcoma.
Authors: Ulmasov Firdavs , Djuraev Mirjalol , Yusupbekov Abrorjon , Egamberdiev Dilshod Makhmudovich
Published: September 2018

e22537

Background: to improve the efficiency of the treatment of retroperitoneal fibrosarcomas by developing complex approach with using neoadjuvant endoarterial chemotherapy, surgical intervention followed by radiotherapy. Methods: For material of the study we analyzed treatment results of 34 patients with retroperitoneal giant fibrosarcoma. Depending on the method of treatment patients were divided into 2 groups: 1 main group n = 16, underwent neoadjuvant chemotherapy + surgery + radiotherapy 2 control group n = 18 who underwent surgery + adjuvant chemotherapy. There was performed endoarterial catheter into the close blood vessels feeding the tumor at the preoperative period. Chemotherapy was conducted 3 cycles by CAP regimen. Drugs were administered endoarterially with 48 hours infusion. After receiving chemotherapy for all patients there was performed radical surgery. Postoperatively patients received radiotherapy to the tumor bed with single dose 1.8-2 Gr and up to total dose - 60 Gr. Results: Therewere selected the following criteria: - The effect of chemotherapy on tumor size, extension and traumatizing of surgical treatment, and the duration of relapse-free and metastases-free period. - The effectiveness of postoperative radiotherapy for the duration of disease-free period. Chemotherapy in the preoperative period, led to partial regression of tumors in 9 (58.3%) patients. In 4 (25.0%) patients the process was stabilized, in 3 (16.7% 5 + 15%) observed progression of the process. Study of patients’ survival, which is the main determining factor showed an increase of survival in the main group up to 18.2 +0.4 months, and in the control group 12.4 +0.3 months. (P < 0.05). Recurrence free survival in the main group was 10,2 +0,4 months, and in the control group 7,3 +0,3months (P < 0.05), metastases-free survival consisted 11,4+0,5 months and 9,1+0,4 month, respectively.(P < 0.05). Conclusions: Complex approach in the treatment of retroperitoneal fibrosarcomas significantly increased the overall survival and as well as the relapse-free and metastases-free survival. Using adjuvant continuously endoarterial chemotherapy gives partial effect in 58.3% cases, and thereby leads to more favorable conditions for surgery.
Comparative analysis of cell cycle parameters in primary and recurrent soft tissue sarcomas.
Authors: Novikova Inna Arnoldovna , Nepomnyashchaya Evgeniya M. , Aliev Timur , Zlatnik Elena Yurievna , Selyutina Olesya N. , Ulianova Elena Petrovna , Vashchenko Larisa Nikolaevna , Ausheva Tatiana Valeryevna Ausheva , Andreyko Elena A.
Published: September 2018

e22538

Background: The purpose of the study was to determine DNA content and distribution of cells in cell cycle phases by flow cytometry in patients with primary and recurrent soft tissue sarcomas. Methods: 60 patients with soft tissue sarcomas (STS) were recruited: 30 with primary tumors and 30 with recurrent ones. Mean age of patients with primary STS was 56±5.4 years, with recurrent STS – 55±6.7 years. DNA content was determined using the BD Facs Cantoo II flow cytometer with CycleTEST PLUS DNA Reagent Kit (Becton Dickinson). The data were processed using ModFit LT program. Results: Comparative analysis of the cell cycle kinetics showed an increase in the percentage of cells in G2+M phase by 2 times in diploid and by 2.1 times in aneuploid recurrent tumors in comparison with primary ones (1.8±0.5% vs. 0.9±0.1% for diploid tumors; 5.4±2.2% vs. 2.6±0.7% for aneuploid tumors). An increase in the percentage of aneuploid tumors was found in recurrent G2 and G3 tumors (from 50% in primary to 66.7% in recurrent G2 tumors and from 63.25% in primary to 85% in recurrent G3 tumors). Mean content of aneuploid cells in recurrent G2 tumors was 2.2 times higher (p≤0.05), while the differences in primary and recurrent G3 tumors were not significant. The percentage of aneuploid tumors depended on the disease stage and increased in stages IIb and III in recurrent tumors, compared to primary ones (from 37.5% to 71.4% in recurrent st. IIb; and from 65% in primary st. III to 72.7% in recurrences) (p≤0.05). Conclusions: DNA analysis by flow cytometry demonstrated a high biologic potential of both primary and recurrent tumors. Some values in the mitotic cycle in recurrent tumors were probably associated with adjuvant therapy, as well as influenced by the coefficient of two parameters – the percentage of cells in G2+M phase and the cell loss factor determining a high malignant potential of these tumors.
GGNBP2 to suppress tumor growth and cancer stem cell load of triple negative breast cancer by controlling STAT3 pathway.
Authors: Zhang Jin , Hu Yunhui , Lei Zhenmin
Published: September 2018

e23003

Background: Gametogenetin binding protein 2 (GGNBP2) is encoded in human chromosome 17q12-q23, a region known as a breast and ovarian cancer susceptibility locus. GGNBP2 has a single C2H2 zinc finger and a consensus LxxLL nuclear receptor (NR) binding motif. We have reported that GGNBP2 suppresses ERα-positive breast tumorigenesis by acting as a nuclear receptor co-repressor to restrain ERα activity. However, the detailed molecular mechanisms of GGNBP2 and its role in triple negative breast cancer (TNBC) remain largely unclear. Methods: A human breast cancer tissue array containing 138 human breast tumor tissues were utilized to examine GGNBP2 expression in breast cancer samples by IHC. To address the potential anti-breast tumor activity of GGNBP2 in vitro, we expressed exogenous GGNBP2 in TNBC cells, including MDA-MB-231 and Cal51 cell lines. Cell proliferation and cell cycle were assessed by cell growth curve/EdU assays and flow cytometry after propidium Iodide staining. Apoptosis was determined by flow cytometry after annexin V staining, by caspase 3/7 and caspase 9 activity assays. Cancer stem cell properties were determined by expression of CD44/CD24/ALDH1 markers. The levels of phosphorylated STAT3 and total STAT3 were determined by western blot. Quantitative PCR and Western blot were carried out to evaluate the effects of GGNBP2 overexpression on STAT3 target genes, CCND1, Mcl-1, survivin, bax and bim expression. Results: GGNBP2 expression is down-regulated in TNBC cells and patient tumors. GGNBP2 down-regulated expression is associated with poor patient survival. Overexpression of GGNBP2 significanly induces cell cycle G0/G1 phase arrest and apoptosis in TNBC cell lines. Expression of cancer stem cell markers also decreased in GGNBP2-overexpressed TNBC cells. GGNBP2 reduces the expression levels of CCND1, Mcl-1 and survivin, promotes the expression levels of bax and bim proteins. Importantly, overexpression of GGNBP2 inhibits STAT3 phosphorylation and STAT3 downstream target gene expression, including CCND1, Mcl-1 and survivin. Conclusions: GGNBP2 serves as a critical nuclear negative regulator of STAT3-mediated gene expression and tumorigenesis in TNBC.
NIDOGEN1 as a novel regulator of endothelial control over breast cancer invasion and metastasis.
Authors: Buess Martin , Ferraro Daniela A. , Patella Francesca , Zanivan Sara , Christofori Gerhard
Published: September 2018

e23005

Background: The microenvironment is a central regulator of cancer biology. While the contribution of fibroblasts has been largely studied, the role of endothelial cells as regulators of cancer cell behavior is still poorly understood. As in a diverse spectrum of physiological processes in normal tissue, endothelial cells may exert a similar regulatory control in cancer progression and metastasis. Methods: To characterize the functional effects of endothelial-cancer interaction we focused on an in vitro co-culture model. Results: Co-culturing human umbilical venous endothelial cells (HUVEC) with SKBR-3 breast cancer cells induced morphological changes with epithelial-mesenchymal transition traits (EMT) and a significantly increased migratory and invasive potential. This activity leading to an elongated phenotype, expression of mesenchymal markers and pro-migratory gene sets in SKBR-3 was contained in HUVEC conditioned medium. The pro-migratory effect on SKBR-3 was significantly more pronounced when the supernatant was obtained from a sub-confluent and highly proliferative endothelial cell culture than from a confluent and resting endothelial cell layer. To identify the secreted regulatory molecules, we analyzed the supernatant of sub-confluent and confluent endothelial cells by quantitative MS proteomics (SILAC analysis). Eight candidate proteins significantly more secreted in conditioned medium from confluent HUVEC represented potential inhibitors of migration. Among them NIDOGEN1 was found to be necessary and sufficient for the inhibition of EMT and migration in SKBR-3. Stimulation of SKBR-3 with supernatant from sub-confluent HUVEC increased p-STAT3 levels in SKBR-3. Silencing nidogen1 in confluent HUVEC re-activated phosphorylation of STAT3 indicating that NIDOGEN1 inhibits the promigratory STAT3 pathway. The STAT3 pathway and migration were also inhibited by overexpression of nidogen1 in MDA-MB-231 LM2 cells.When injected in the mammary fat pad of nude mice these cells formed significantly less lung metastases than controls (p < 0.01). Conclusions: We identified NIDOGEN1 as a novel regulator of endothelial control over cancer cell migration, invasion and metastasis.
miR-221 to modulate tumor growth in vivo and as a regulator of TGF.
Authors: Spahn Martin , Zoni Eugenio , Krebs Markus , Herreiner Philip , Kalogirou Charis , Chen Lanpeng , Snaar-Jagalska Ewa , Thalmann George N. , Kruithof-de Julio Marianna , Riedmiller Hubertus , Kneitz Burkhard
Published: September 2018

e23008

Background: Despite the advances in cancer therapy, when Prostate Cancer (PCa) progress to castration resistant phase, patients develop incurable bone metastases. Understanding the processes that regulate homing and survival of metastatic cancer cells in the bone is crucial for the identification of new therapies. TGF-β signaling plays a major role in bone remodeling and according to the “vicious cycle hypothesis” is a master regulator of maintenance of prostate cancer cells in lytic bone lesions. microRNAs (miRs) are a class of small non-coding RNAs that regulates many biological process. miR-221 expression has been previously associated with prostate cancer progression. Here we studied the effect of miR-221 on TGF-β signaling and the impact of miR-221 on tumor growth in vivo Methods: miR-221 was overexpressed in PC3 and RWPE-1 cells and PMEPA was monitored by RT-qPCR and western blot. Expression of miR-221 in PC3 and RWPE-1 cells was assessed by RT-qPCR. Luciferase assay was used to investigate the interaction between miR-221 and PMEPA1. For zebrafish experiment, fluorescently labelled PC-3M-Pro4 cells overexpressing miR-221 were injected in the Duct of Cuvier (DC) of zebrafish embryos. Results: miR-221 overexpression resulted in decreased PMEPA1 mRNA and protein in PC-3 cells. Luciferase reporter assay indicated that miR-221 can directly interact with PMEPA1 3’ UTR. We show an enhancement of the proliferative effect of TGF-β in PC-3 cells, following miR-221 overexpression. In conclusion, we observed increased Smad2 activation upon TGF-β treatment in miR-221 overexpressing PC-3 cells. Inoculation of PC-3M-Pro4 cells overexpressing miR-221 in the DC of zebrafish embryos resulted reduced tumor burden compared to control. Finally, we observed inverse correlation between miR-221 and PMEPA1 expression in normal vs. tumor tissue collected from PCa patients. Conclusions: Our results indicate that miR-221 is a regulator of TGF-β signaling via modulation of PMEPA1 and miR-221 overexpression can reduce tumor growth in vivo.
Low T3 syndrome with hyperthyrotropinemia and early detection of liver metastasis.
Authors: Kaplieva Irina V. , Frantsiyants Elena Mikhaylovna , Trepitaki Lidia K. , Pogorelova Yulia A. , Bandovkina Valeria , Tkalya Lyudmila Dmitriyevna , Islamova Elena F. , Cherkes M. A. , Kit Oleg I.
Published: September 2018

e23010

Background: Numerous clinical studies have shown an association between changing thyroid gland (TG) function and the development of tumors. The purpose of the study was to analyze the dynamics of thyroid hormones in the pituitary gland, TG and blood serum (BS) during liver metastasis in order to create thyroid profile of metastasis and to detect thyroid markers of metastasis in BS. Methods: The experiment included 44 white male rats weighing 180-250 g. Sarcoma 45 was transplanted intrasplenically. Levels of thyroid stimulating hormone (TSH) were studied in the pituitary gland, TG, BS; the amount of free (fT4) and total (T4) thyroxine and free (fT3) and total (T3) triiodothyronine in TG and BS were determined by radioimmunoassay (Immunotech, Czech Republic; Arian analyzer, Russia). Results: Exertion of the pituitary gland and overproduction of TSH were observed since the early days of the tumor development, later accompanied by TG hypofunction. Quantitative changes of thyroid hormones in organs did not coincide with their dynamics in BS. The first diagnostic signs of experimental liver metastasis, in the absence of formed metastases in the body, were hyperthyrotropinemia and a tendency to fT3 decrease in BS. Already formed liver metastases were characterized by a marked low T3 syndrome which transformed into a more severe lowT3/lowT4 syndrome during the secondary metastasis. Conclusions: The development of liver metastases is accompanied by the exertion and imbalance of the thyroid system function. Analysis of dynamics of the thyroid axis hormones in BS allows predicting the appearance of liver metastasis and identification of “the point of no return” in the development of the pathology that leads to secondary metastasis and irreversible progression of the disease.
A novel role for 17-Β estradiol and testosterone in osteosarcoma tumorigenesis and metastasis.
Authors: Lichterman Jake N. , Post Robert , Menken Luke , Saad Bishoy , Delgado Justin , Sall Lauren , Karr Matthew , Kozlowski PIotr , Degenhardt Kurt
Published: September 2018

e23012

Background: Pulmonary metastases still remains the leading cause of mortality in osteosarcoma, with a 5-year survival rate of less than 30%. 17-β estradiol and testosterone are known to promote tumorigenesis and metastasis in other cancers, but its role in osteosarcoma tumorigenesis and metastasis is unclear. An understanding of the mechanism that these hormones have in osteosarcoma tumorigenesis and metastasis will lead to new therapeutic strategies using currently available targeted therapies used in breast and prostate cancer. Methods: We began by assessing the in vitro characteristics of two human osteosarcoma cell lines treated with 17-β estradiol or testosterone via proliferation and invasion assays. For the in vivo experiments, NCR nu/nu mice were injected with two luciferase-tagged human osteosarcoma cell lines and treated with 17-β estradiol and testosterone slow-release pellets. We first injected the mice subcutaneously and monitored tumor growth via caliper measurements. In the second experiment we injected the cell lines intratibially and monitored primary tumor growth in the bone and monitored metastasis formation in the lungs via IVIS bioluminescent imaging. Quantification of luciferase signal (photons/second) in the lungs was performed via the Xenogen Living Image software. H&E and immunohistochemistry for luciferase expression was performed on lung tissue to quantify tumor burden in the lungs. Statistical analysis was performed using Graphpad Prism. Results: Treatment with 17-β estradiol and testosterone increased cell proliferation and invasion in a dose-dependent manner. We observed increased tumor growth with both 17-β estradiol and testosterone in the subcutaneous model. In the intratibial experiment, testosterone increases primary tumor growth, but has no effect on metastasis. 17-β estradiol decreases primary tumor growth, but increases metastasis in a cell line independent manner. Conclusions: This study demonstrates a novel role for the puberty-related sex steroid hormones, 17-β estradiol and testosterone, in osteosarcoma tumor growth and metastasis. Uncovering the mechanism behind this phenomenon will uncover new therapeutic options for this devastating disease.
Oncogenic microRNAs to predict relapse in early breast cancer patients.
Authors: Sochor Marek , Basova Petra , Pesta Michal , Bartos Jiri , Stopka Tomas
Published: September 2018

e23021

Background: Early breast cancer is a frequent female disease with different outcomes. New approaches are needed in order to improve its prognosis. MicroRNAs (miRs) are modulators of gene expression and act as oncogenes and function to inhibit tumor suppressors and to promote metastasizing. Monitoring of miRs could be of benefit to the prognosis of EBC patients. Methods: We prospectively collected sera from 133 EBC patients (follow-up 53,25 months) in 3 time points (before I and after surgery II, after therapy III). Patients were stratified into high and low-risk groups according to HR, HER2, Ki-67, grade, LN. For RNA isolation serum was used followed by RT-qPCR and was applied longitudinal multivariate data analysis. Aim of the project was to determine serum expression of miR-155, miR-19a, miR-181b, miR-24 in 3 time points, to find difference in expressions between high and low-risk groups, to find association between miRs and clinical/pathological risk factors and associations in miRs expression and prognosis of EBC. Results: EBC patients significantly over-express miRs in time point I. In time point II the levels of miR-155, miR-181b, miR-24 significantly decreased ( p< 0.05). miR-19a decreased in time point III ( p= 0.00869). Levels of miR-155, miR-19a, miR-181b, miR-24 are significantly more abundant in high-risk in comparison to low-risk patients ( p< 0.05) and decrease upon therapy. The multivariate GEE model revealed that miR-155, miR-24 were predictive of the relapse ( p= 0.025 and 0.041). miR-19a, miR-181b are insignificant with respect to the relapse ( p> 0.05). Triple-negativity, HER2+, grade III, LN+ have no effect on the probability of relapse ( p> 0.05) when miRs are simultaneously taken into an account. The only risk factor that makes the prediction of relapse more precise is Ki-67 > 20% ( p= 0.013 in case of miR-155 and p= 0.023 in case of miR-24). Conclusions: OncomiRs are significantly more abundant in EBC patients at diagnosis and decline after therapy. Differences in miR levels reflect EBC risk groups. The data shows that miR-155 and miR-24 enable monitoring of EBC and predict relapse independently of clinical/pathological risk factors. Combining the miR levels with Ki-67 expression further specifies the relapse probability.
Difference in appearance of KRAS mutated circulating tumor DNA in patients with metastatic colorectal cancer during treatments.
Authors: Takayama Yuji , Suzuki Koichi , Ichida Kosuke , Fukui Taro , Watanabe Fumiaki , Kakizawa Nao , Muto Yuta , Kikugawa Rina , Hasegawa Fumi , Tsujinaka Shingo , Noda Hiroshi , Miyakura Yasuyuki , Konishi Fumio , Rikiyama Toshiki
Published: September 2018

e23025

Background: Emergence of KRAS mutation in blood is observed in colorectal cancer patients who undergo chemotherapy, but its clinical significance is not well known. In this study, we focused on the difference in appearance of KRAS mutated circulating tumor DNA (MctDNA) and elucidated its association with treatments. Methods: Four hundred and fifty-one plasma samples were collected prospectively from 85 patients (pts) who underwent chemotherapy due to metastatic colorectal cancer in 2014 - 2016. Seven types of KRAS mutation in MctDNA were detected by droplet digital PCR creating oil droplets. To exclude false positive detection, mutation was validated. MctDNA amplified in oil droplets was selectively sorted by On-chip sorting system and mutation was determined by Sanger sequencing. Results: KRAS assessment in tumor tissues showed 29 pts with KRAS mutation (MT), 56 pts without KRAS mutation (WT). Among 29 pts with MT, KRAS assessment in plasma displayed 23 pts with MctDNA and 6 pts without MctDNA. The type of mutation in MctDNA was consistent with that detected in tumor tissues, indicating mutual exclusivity in KRAS mutation was confirmed. In 56 pts with WT, 28 pts showed MctDNA during treatments. Difference in appearance of MctDNA was recognized in several treatments. Gradual increase in detection of MctDNA was observed with anti-EGFR antibody, resulted in treatment resistance. Transient spike elevation was frequently seen in TAS-102, which associated with drug response. No specific appearance was recognized during treatments with other drugs including anti-VEGF antibody. MctDNA in oil droplets were successfully sorted even if a few droplets were targeted, and mutation was confirmed. Conclusions: Difference in appearance of MctDNA may associate with treatment response in patients with metastatic colorectal cancer during treatments. [Table: see text]
Circulating microRNAs (miRNAs) as predictors of relapse in early breast cancer (BC).
Authors: Agelaki Sofia , Papadaki Chara , Stratigos Michalis , Spiliotaki Maria , Markakis Georgios , Mastrostamatis Georgios , Mavroudis Dimitrios
Published: September 2018

e23030

Background: Disease relapse is usually a lethal development following treatment of early BC. Recurrence could arise from a state of tumour dormancy during which there is growth restriction of undetectable micrometastases. The expression of dormancy and metastasis related miRNAs was evaluated in the plasma of patients (pts) with early BC obtained before adjuvant therapy in order to discover novel biomarkers for the prediction of relapse. Methods: PlasmamiR-21, miR-23b, miR-190, miR-200b and miR-200c expression was assessed by qRT-PCR in 133 pts with early BC (non-relapsed, n = 84; relapsed, n = 49). Expression was classified as high or low according to the median values. Results: No correlation was observed between the expression of miRNAs and the clinicopathological characteristics of pts. After a median follow-up of 94 mo, median Disease Free Interval (DFI) was significantly lower in pts with high compared to low miR-21 [105.03 mo vs not reached (NR); p= 0.001], miR-200c (105.2 mo vs NR; p= 0.007), or both miR-21 and miR-200c expression (81.37 mo vs NR; p= 0.001). miR-21-high was also associated with decreased median Overall Survival (OS; p= 0.041). In multivariate analysis the number of infiltrated axillary lymph nodes (N3 vs N0-N2, HR: 2.86; p= 0.004) and miR-21 expression (high vs low, HR: 2.824; p= 0.001) were independent negative prognostic factors for DFI, whereas negative hormone receptor status (HR: 3.062; p= 0.024) and miR-21 high (HR: 3.545; p= 0.029) independently predicted for worse OS. Moreover, miR-21 expression was higher in pts with early relapse (defined as relapse at ≤ 3 yrs) compared to non-relapsed at 5 yrs ( p= 0.032). In addition, higher miR-21 ( p= 0.038), miR-23b ( p= 0.039), miR-200b ( p= 0.027) and miR-200c ( p< 0.001) levels were observed in pts with late relapse (at ≥ 5 yrs) compared to those without relapse. Conclusions: Differential expression levels of metastasis and/or dormancy related circulating miRNAs are detectable before adjuvant therapy in pts with early BC presenting subsequent relapse compared to non-relapsed pts. Circulating miRNAs can predict for early or late recurrence years before clinical detection of metastases. These results merit prospective validation in an independent patient cohort.
Performance of Biocept's sample collection for tumor cell analysis.
Authors: Singh Veena M. , Sales Edgar V. , Huynh Lan , Vibat Cecile Rose T. , Barón Anna E , Feser William , Malinowski Heather , Merrick Daniel , van Bokhoven Adrie , Malkoski Stephen , Miller York E , Wolf Holly J , Varella-Garcia Marileila
Published: September 2018

e23036

Background: Liquid biopsy is a minimally invasive and cost effective way to assess cancer biomarkers without the risk of surgical biopsy complications. Circulating tumor cell (CTC) analysis from body fluids can provide critical information towards early detection, prognosis and treatment decisions. Accurate CTC evaluations require optimal cell preservation. Cell lysis, DNA degradation, or membrane alterations compromise CTC analyses and accurate diagnoses. This work compares Biocept’s proprietary CEE-Sure BCT and Saccomanno's Cytology Fixative largely used for sputum collection. Methods: One million BT474 (HER2 amplified) or H3112 (ALK re-arranged) cells were spiked into 500 µl medium; 500 µl of CEE-Sure or Saccomanno fixative was added. Tubes were stored at 4°C for 1 day, 1 week, or 1 month. Cells were centrifuged, resuspended, and counted (Celigo). Around 150 cells in 15 µl of RPMI medium were flowed into Biocept's microfludic system for cell capture; recovery (%) was calculated. Captured cells were subjected to fluorescent in situ hybridization (FISH) analyses for qualitative signal evaluation. Results: As similar results were observed for both cell lines and all time points, combined data will be shown. Median cell recovery after CEE-Sure incubation was 14.1% (range 1.7–44%, n = 12) vs 5.4% (range 0.07–26.9%, n = 12) in Saccomanno's fixative. Median cell capture of ~150 cells fed into Biocept’s microchannel was 96% (range 72-98%) for CEE-Sure vs 82% (range 21-96%) for Saccomanno. Paired t-tests showed significant differences for both recovery and capture. FISH signals from CEE-Sure samples were qualitatively rated Fair to Good, while Saccomanno samples had Poor to Fair, grainy, non-specific signals. Conclusions: This preliminary work shows consistently higher cell recovery, better cell membrane maintenance, and higher quality FISH signals for samples stored in Biocept's CEE-Sure vs Saccomano’s fixative. With liquid biopsy testing gaining rapid traction, maximal cell stability during the transport and storage are crucial. Additional fixative comparison is ongoing in various patient specimen types. These results support expansion of molecular analyses in sputum samples enriched for lung epithelial cells.
Clinical significance of monitoring KRAS in the blood of pancreatic cancer patients.
Authors: Watanabe Fumiaki , Suzuki Koichi , Ichida Kosuke , Takayama Yuji , Fukui Taro , Kakizawa Nao , Endo Yuhei , Kaneda Yuji , Noda Hiroshi , Rikiyama Toshiki
Published: September 2018

e23042

Background: KRAS monitoring provides valuable information for early diagnosis and prediction of treatment outcome in colorectal cancer. KRAS mutation is observed in only half of colon cancer patients, whereas it is detected in 90% of pancreatic cancer patients. Therefore, investigating tumor DNA in blood by KRAS monitoring may be even more valuable in pancreatic cancer patients. In this study, we elucidated the clinical significance of KRAS monitoring in pancreatic cancer patients during treatment. Methods: KRAS in tumor tissues was analyzed for mutations by Scorpion ARMS or RASKET in 38 patients with pancreatic tumors. KRAS in blood was analyzed for mutations (G12D, G12V, G12C, G12A, G12S, G12R, G13D, Q61L, and Q61H) using droplet digital polymerase chain reaction in 23 patients who underwent chemotherapy and who had KRAS mutation in their tissues. Results: KRAS mutation in tumor tissues was detected in 27 of 38 patients (71.1%). These 27 patients showed significantly poorer prognosis (24.4 months median overall survival time [MST]) than the six patients without mutation (p = 0.04), who were all alive. Patients with mutation of 12D in tumor tissues showed better prognosis than patients with other types of mutation; thus, mutation of 12D may have a different biological effect to other mutations. Monitoring of KRAS in blood revealed KRAS mutation in 14 of 23 patients (60.9%). These 14 patients exhibited significantly poorer treatment outcomes than patients without mutation. The MST of patients with KRAS mutation in their blood was significantly shorter than that of patients without KRAS mutation (17.7 months vs. 44.3 months, p = 0.03). Conclusions: KRAS mutation in blood could be a valuable predictive and prognostic biomarker in pancreatic cancer patients who undergo chemotherapy. Additionally, assessment of KRAS in tumor tissues may provide information about individual tumor biology.
Circulating tumor cell (CTC) as a significant prognostic factor in resected primary lung cancer.
Authors: Yoneda Kazue , Hashimoto Masaki , Takuwa Teruhisa , Matsumoto Seiji , Okumura Yoshitomo , Hasegawa Seiki , Tanaka Fumihiro
Published: September 2018

e23044

Background: Circulating tumor cell (CTC) is a potentially useful marker in early diagnosis and monitoring therapeutic effects for patients with malignant tumors, but clinical significance of CTC in primary lung cancer remains unclear. We previously showed that CTC was a useful surrogate marker of distant metastasis in primary lung cancer (Clin Cancer Res 2009). In this study, we evaluated the prognostic value among completely resected patients after long-term follow-up. Methods: A total of 94 patients (median age, 68 years; 30 females and 64 males) who underwent complete resection for primary lung cancer (4 with small cell and 90 with non-small cell) were prospectively evaluated. At the time of enrollment into the study, 7.5mL of peripheral blood was sampled from each patient, and an EpCAM-based detection system (CellSearch) was used for detection of CTC. CTC was detected in 16 patients (CTC-positive, 14.9%). Results: CTC-positivity was significantly associated with a poor recurrence-free survival (5-year recurrence-free survival rate, 40% versus 72%; p<0.01) (Table 1), which was confirmed by a multivariate analysis (hazard ratio, 2.57 [95% CI, 1.26-5.26]; P=0.010). CTC-positivity was also associated with a poor overall survival (5-year recurrence-free survival rate, 62% versus 84%; p<0.05) (Table 1), which was confirmed by a multivariate analysis (hazard ratio, 2.76 [95% CI, 1.14-6.71]; P=0.025). Conclusions: CTC-positivity was associated with poor recurrence-free survival and poor overall survival in resected lung cancer. [Table: see text]
cfDNA mutation frequency in early stage breast cancer.
Authors: Kota Karthik Joshua , Gyanchandani Rekha , Rosenzweig Margaret Quinn , Brufsky Adam , Huggins-Puhalla Shannon Leigh , Miller Lori , Oesterreich Steffi , Lee Adrian V.
Published: September 2018

e23047

Background: Though 93% of patient present with early stage breast cancer (EBC), metastatic recurrence is expected in 7-13%. To prevent estrogen receptor (ER) positive disease recurrence, endocrine therapy (ET) is given after local disease. ET guidelines recently extended therapy —specifically tamoxifen—from 5 years to 10 based on ATLAS and aTTom. This pilot study used blood samples from women 6 months post-ET to determine cell free DNA (cfDNA) mutation prevalence in disease-free patients Methods: Patients with EBC and continued follow-up after ET were recruited from the Magee Women’s Breast Cancer Clinic. Inclusion criteria were EBC, ER+, and completion of ET > 6 months prior to visit; the exclusion criterion was active disease. Patients gave 2 blood samples, placed in Streck and EDTA tubes and processed at 2 laboratory sites for cfDNA. Blood from patients with metastatic breast cancer (MBC) served as controls for mutation detection. cfDNA was amplified for ESR1 and PIK3CA genes. Targeted amplifications underwent digital droplet PCR to identify mutations: 4 for ESR1 (D538G, Y537C/N/S), 2 for PIK3CA (E545K, H1047R) Results: Ten EBC patients > 6 months post-ET (post-EBC; 2/5/3 of stage I/II/III) and 10 MBC patients gave samples. cfDNA yield between plasma isolated from EDTA and Streck tubes (including mutation allele frequencies) was not significantly different (p > 0.05); cfDNA yield from patients with MBC was > 2X higher than post-EBC (p < 0.001). MBC cfDNA had 2 monoclonal and 1 polyclonal (2 different) mutations in ESR1, while post-EBC cfDNA had none Conclusions: This pilot study shows cfDNA can be consistently isolated from Streck or EDTA-processed blood from patients with MBC and post-EBC; however, cfDNA levels are significantly higher in MBC. There were no mutations (ESR1 or PIK3CA) in post-EBC samples, though ESR1 mutations were found in MBC. Further studies are needed to determine if mutations in cfDNA can be found in patients without evidence of disease
Salivary miRNA-210 involvement in medication-related osteonecrosis of the jaws.
Authors: Picone Antonio , Marcianó Antonia , Dimarco Rosanna , Miano Eleonora , Zacchia Alessandra , Sanò Maria Vita , La Fauci Alessia , Chiarenza Maurizio , Ali Marco , Di Leo Maria Grazia , Aiello Rosa Anna , Taibi Eleonora , Fallica Giuseppa , Caruso Michele
Published: September 2018

e23052

Background: Improved survival rates from cancer have increased the need to understand the health related problems of cancer treatment. Persistent oral health related problems significantly affect patient's quality of life. Medication-related osteonecrosis of the jaws (MR-ONJ) is an oral complication of antiresorptive medications, denosumab and other medications that target angiogenesis for the management of metastatic bone cancer. The purpose of this pilot study, is to investigate, for the first time, microRNAs expression in MR-ONJ. Recent studies reported the miRNA-210 (a specific angiogenic-related miRNA) demethylation in the Osteonecrosis of the Femoral Head. We hypothesized that MiRNA-210 could also be involved in MR-ONJ development. Primary objective of the study is to investigate the role of miRNA-210 in MR-ONJ pathogenesis. Secondary objective is to investigate the feasibility of salivary miRNAs detection as potential non invasive biomarkers for MR-ONJ. Methods: A case-and-control study has been designed. Inclusion criteria are malignant tumors with ongoing treatment or previous history of anti-resorptive or anti-angiogenic medications. The predictor variable is the presence or absence of MR-ONJ. Oral examination and dental x-ray examinations suggestive of MR-ONJ will be performed. Saliva sample will be collected according to the protocol by Navazesh. The methylation status of miRNA-210 will be investigated using quantitative PCR (Q-PCR). Values will be compared between case and control group. Results: The main expected results are to enroll approximatively twenty patients in case group and sixty patients in control group within a one year recruitment period, validate that saliva is a medium for the identification of biomarkers associated with MR-ONJ and confirm the hypothesis that miRNA-210 is involved in MR-ONJ development. Conclusions: Results acquired within this project would represent a contribute to the understanding of the molecular mechanisms driving to the development of MR-ONJ lesions. MiRNA-210 demethylation could serve as a potential therapeutic target for the treatment of MR-ONJ through stimulation of angiogenesis.