bluuebunny/crossref_metadata_2025 huggingface dataset:
Identification and validation of stromal immunotype predict survival and benefit from neoadjuvant chemotherapy in patients of breast cancer.
Authors: Ji Fei , Yang Ci-Qiu , Zhang Liulu , Yang Mei , Li Jieqing , Gao Hong-Fei , Zhu Teng , Cheng Min-Yi , Wang Kun
Published: May 2020

e15261

Background: Immune infiltration of breast cancer is associated with clinical outcome. A growing number of research suggests the diversity of functionally distinct cell types that make up the immune response. The aim of this study was to determine whether differences in the cellular composition of the immune infiltrate in breast cancer influence survival and treatment response, and construct the stromal immunotype which could improve prediction of neoadjuvant therapy and survival. Methods: A total of 1502 ER negative breast cancers from TCGA and METABRIC cohort were used to infer the proportions of 22 subsets of immune cells, Another 200 ER negative breast cancer patients from Guangdong Provincial People’s Hospital in the validation cohort were also included in the study. Immune cell infiltration was evaluated by immunohistochemical staining or CIBERSORT method, Five immune features were selected out of 22 immune features to construct immunotype based on LASSO Cox regression model. Results: Of the cell subsets investigated, CD8+ T cells (p < 0.001), CD4+ T cells (p = 0.002), B cells (p = 0.003), M1 macrophages(p = 0.006) were associated with favourable outcome. T regulatory cells(p = 0.005) emerged as the most strongly associated with poor outcome. T regulatory cells were associated with pathological complete response to neoadjuvant chemotherapy (p = 0.012). Using the LASSO model, we classified ER negative breast cancer patients into stromal immunotype A subgroup (CD8+Thigh CD4+Thigh B cellhigh M1 macrophagehigh Treglow) and stromal immunotype B subgroup (CD8+Tlow CD4+Tlow B celllow M1 macrophagelow Treg high). Significant differences were found between immunotype A and immunotype B in the combined cohort with 10-year overall survival (66.2% vs. 49.8%; P < 0.001) and 10-year disease-free survival (63.8% vs. 44.4%; P < 0.001). Stromal immunotype was revealed to be an independent prognostic indicator in multivariate analysis in all cohorts separately, and also showed to be related to pCR in neoadjuvant chemotherapy. Finally, stromal immunotype A showed higher immune checkpoint molecules (PD-L1, PD-1, CTLA-4) expression and three important cytokines expression profiles (IL-2, INF-γ and TGF-β). Conclusions: The stromal immunotypes could predict survival and recurrence of ER negative breast cancer patients effectively. Furthermore, the immunotypes might be a practical predictive tool for immunotherapy.
Oncolytic activity of a new group of non-classified rotavirus of Reoviridae family on human epidermoid carcinoma A431 growth.
Authors: Kolpakov Sergey A. , Kolpakova Elena P. , Zlatnik Elena Yu. , Nepomnyashchaya Evgeniya M. , Shulgina Oksana G. , Sitkovskaya Anastasia O. , Kit Oleg I.
Published: May 2020

e15272

Background: Significant increase of malignant tumors` incidence all over the world in spite of vast arsenal of anticancer drugs and their combinations challenges the oncologists to develop new approaches to tumor treatment. Application of oncolytic virus is considered to be rather promising. Here we studied two strains of the new group of non-classified rotavirus of Reoviridae family (RVK): 100 and 228 ( http://jbks.ru/archive/issue-10/article-6 ). RVK are attenuated non-pathogenic virus growing on pig embryo kidney cell culture with concentration 5·109 per 1 ml. The aim of the study was to assess the effect of two RVK strains (100 and 228) on the growth of human epidermoid carcinoma A431 transplanted to nude mice in vivo. Methods: Mice Balb/cNude 22-24 g body weight (n = 13) were injected with 4х106 cells of human epidermoid carcinoma A431 subcutaneously in SPF-area of vivarium. After formation of palpable tumors (1 week after transplantation) administration of alive RVK strains 100 and 228 was performed. The 3rd group was the control one and received 0.85% NaCl. The injections were performed once a week 0.3 ml. Tumor growth rate and its` volume were measured. 1 week after the completion of the course mice were sacrificed, tumors were weighted and their morphology was studied. Results: In mice injected with RVK tumor growth inhibition developed in early date after the injection and was statistically significant within 30 - 40 days of monitoring only after the administration of strain 100. In these animals tumor weight was 2.6 times lower than in control mice (6.3±3.0 и 16.6±2.3 g respectively, р < 0.05), in mice having received strain 228 it was 1.7 times less tan in controls. Layers of high-grade sqamous carcinoma cells without keratinization with inflammatory and necrotic foci were observed in tumors of control mice while in tumors inhibited by RVK dystrophic changes and fragmentation, inflammation in places severe and necrotic foci were seen. Implication of the strain 100 resulted in 2-3 time reduction of tumor size, decrease of tumor cells` layers, mononuclear infiltration. Conclusions: In the in vivo model of human epidermoid carcinoma A431 transplanted to nude mice the inhibition of tumor growth under RVK administration was established. Our results confirm the oncolytic activity in RVK, particularly in strain 100.
VEGF-A polymorphisms affect the pharmacologic effect of bevacizumab on VEGF-A levels in patients with metastatic colorectal cancer.
Authors: Papachristos Apostolos , Kemos Polychronis , Kalofonos Haralabos , Sivolapenko Gregory
Published: May 2020

e15505

Background: Bevacizumab treatment presents large interpatient variability in efficacy. Mutations in VEGF-A rs699947 are associated with longer overall survival and progression free survival in bevacizumab-treated metastatic colorectal cancer (mCRC) patients. The aim of the present study was to investigate correlations between VEGF-A polymorphisms and VEGF-A levels in patients receiving bevacizumab for mCRC. Methods: 46 patients were studied. Blood samples were collected for VEGF-A (rs2010963, 1570360, rs699947) genotyping, and measurement of bevacizumab and VEGF-A levels. The Spearman's rank correlation coefficient was used to study the correlation between VEGF-A and bevacizumab levels. For the multiple analysis, we used linear regression with residual analysis and considering multicollinearity while the effects are reported with 95% confidence intervals. Results: In total 171 samples for VEGF-A and 157 for bevacizumab levels were analyzed. A very strong negative correlation between bevacizumab and VEGF-A levels (coef. = -0.625, p < 0.000001) was noted. Interestingly, VEGF-A rs699947 plays an important role in the model (p = 0.0002, 95% CI 32.5-58.1) and as VEGF-A levels were found to be significantly lower in patients with mutant rs699947 (p < 0.0001, 95% CI 25.2 - 53.8). Conclusions: In conclusion, genetic factors significantly affected the effect of bevacizumab on VEGF-A levels. Significantly lower VEGF levels were measured in carriers of mutant rs699947. These results may explain the favorable clinical outcomes reported in this sub-group of patients. These findings might enable clinicians to distinguish patients, who may benefit more and in a long-term. Finally, can be used to design individualized dosing schemes for bevacizumab.
A rare case of metastatic testicular adult granulosa cell tumor.
Authors: Natoli Guido , Metovic Jasna , Taraglio Stefano , Miraglia Stefania , Misischi Francesca , Delsedime Luisa
Published: May 2020

e15506

Background: Adult granulosa cell tumour (AGCT) is a rare disease that develops in the testis, composed of sex cord cells, accounting for less than 0.5% of all sex cord-stromal neoplasias (WHO 2016). To our knowledge, 48 cases of AGCT of the testis have been published up to date reporting follow-up (Table 1). Infrequently, these tumors can metastasize, usually to retroperitoneal lymph nodes, but also liver, lungs and bones. Methods: In 2015 a 60-year-old Caucasian male with unilateral, painless slowly growing mass underwent to orchioectomy. A diagnosis of malignant ADTC was rendered. After 19 months metestatic spread to lung was hystologically confirmed. After cytotoxic treatment a compassionate use of pazopanib was sterted and the was in complete response today. Results: The patient is still on pazopanib treatment with complete response that is well tolerated and has improved quality of life 55 months after the initial AGCT diagnosis. Conclusions: Our case report could offer an important source of information for clinicians who are currently facing this rare entity. In summary in case of tumor larger than 4-5 cm adjuvant treatment may be considered and, more importantly, systemic treatments, including pazopanib, could be suggested as a treatment apporach to a metastatic AGCT patient. In the future, other reports containg long-term follow up will be of vital importance to understand the best management of patient with AGCT.
Stability and visual compatibility of daunorubicin and cytarabine during Y-site administration with selected drugs.
Authors: Sicard Guillaume , Donnette Mélanie , Pourroy Bertrand , Fanciullino Raphaëlle
Published: May 2020

e15516

Background: Vyxeos is a daunorubicine plus cytarabine liposome indicated in adults patient with newly diagnosed acute myeloid leukemia with myelodysplasia-related changes (AML-MRC) and therapy-related AML (t-AML). The standard protocol is an induction phase on D1, D3 and D5 at 100 mg/m² (expressed in cytarabine) during 90 minutes followed by 2 consolidation cures at 65 mg/m² on D1 and D3. Data provided from manufacturer indicate stability over 4 hours for diluted solution of daunorubicine plus cytarabine. AML-Patients are all receiving complex infusion increasing the risk of Y site incompatibilities. However, there is a lack of data on Y site compatibility of daunorubicine plus cytarabine with others drugs while these phenomena are well described with cytarabine and daunorubicine. The aim of this study was to evaluate stability and container-content interaction over time and visual Y site compatibility with selected drugs. Methods: Physicochemical stability over time was evaluated by Dynamic Light Scattering (DLS) (particle size) and by LC-MS/MS analysis (determination of cytarabine concentration). Daunorubicine plus cytarabine diluted solutions in 0.9% NaCl, final concentration 0.17 mg/mL were stored at 4°C during this study. Container-content interaction was evaluated comparing glass bottle and free-PolyVinylChloride (PVC) bag. Regarding Y site compatibility, a total of 33 drugs were tested. Medication mixtures were analyzed immediately after contact (T=0) and 90 minutes after, stored at room temperature (i.e., 25°C). The particle analyses were performed visually and by DLS. Drugs couple were classified in three categories: compatible, incompatible and variable compatibility when dubious phenomenon was observed. Results: No significative difference was found over time (120h) in term of particle size and chemical stability of cytarabine encapsulated, regardless of the container. This suggests that stability of daunorubicine plus cytarabine could actually be much longer than manufacturer’s recommendations. Most of the tested drugs were compatible. Still, three drugs exhibited visual incompatibilities at T = 0 and T = 90 minutes. However seven drugs presented incompatibilities using LDS. Two combinations presented conflicting data and thus classified as variable compatibility. Conclusions: Polypharmacy is a major concern in AML patients and knowledge and management of daunorubicine plus cytarabine stability and Y site compatibilities are critical to ensure a maximum safety at bedside. At least 10 drugs frequently used in oncology present a variety of incompatibilities with daunorubicine plus cytarabine and therefore should not be co-administered on Y site.
Abscopal effect induced intratumoral injection of chemotherapy drugs with hapten and tumor microstructure changes with protein expression of distal untreated tumors in mice.
Authors: Yu Baofa , Gao Feng , Jing Peng , Liu Jian , Zhang Jian , Li Bian , Yang Huanzhen , Han Yan , Wu Yanfei , Fu Qiang ,   Shandong Baofa Cancer Therapeutic Systems
Published: May 2020

e15518

Background: To observe the abscopal effect by intratumoral injection of chemo drugs with hapten on tumor, and analyze the difference in expression of bilateral tumor proteins and the inhibition of tumor growth on the opposite side. Methods: Balb/c mice 45, H22 ascites were inoculated in left forelimb armpit. When longest diameter of about 4-5mm randomized (day 0); the next day, intratumoral injection different combination drugs, the three days later in the right armpit mice were inoculated inoculated 0.2ml (105/ml) H22 cells, the seven days later of secondary administration, while observing the contralateral sides, especially tumor growth. 15 days after first treatment each group were sacrificed 6 mice, tumor tissue on both sides for immunohistochemistry, quantitative RNA PCR quantitative determination. Results: It showed an upward trend in body weight in mice, 15 days after administration of DNP sustained release drug group and PYM control group of mice were significantly lower than the body weight of the negative control group, the difference between groups (P<0.05); three groups of mice after administration of tumor volume on the left. On the 15th day after the administration, the left and right tumors in DNP group were significantly smaller than those in PYM group and negative control group, and the difference was statistically significant (P < 0.05); the left tumors in PYM group were significantly smaller than those in negative control group, and the difference was statistically significant (P < 0.05). After administration PYM control group’s Col1a1, CD4, Il12aA, TGF, Ela, NFKB, Cox2, CD11c, CD8, TNFa expression was significantly lower than the negative control group and DNP sustained-release drug group; DNP sustained-release drug group’s Col1a1, CD4,Il12aA, TGF, Elastin, NFKB, CD11c, CD8 expression was significantly lower than the negative control group. A large number of cell disintegration phenomenon, observed lymphocytes, leukocytes, the presence of high electron density granular, suggesting that there may be drugs enter into cancer cells. Conclusions: Intratumoral injection of chemotherapy drugs with hapten can exhibit significant anti-tumor effect. In the DNP sustained-release drug group, abscopal effect is more obvious. After the use of PCR quantitative detection of various tumors and fibroplasia, inflammatory factors and immune-related factors have changed significantly to further confirm abscopal effect by intratumoral injection of combination drugs.
Morphology classification of circulating tumor cells could be a predictor of recurrent disease in patients with non-small cell lung cancer after surgery.
Authors: Wu Yun , Niu Yuxu , Lv Fanzhen , Gao Wen , Shen Xiaoyong
Published: May 2020

e15530

Background: CTCs have been widely used in monitoring the efficacy and prognosis of lung cancer. However, CTCs number count alone cannot accurately predict the recurrent disease in patients. In this study, we investigate whether the morphology classification of CTCs could be as a prognostic marker for increased risk of recurrence after surgery. Methods: In this study, 105 lung cancer patients (median age 68y) who underwent surgery were prospectively enrolled in this study. Samples were obtained before, after, and serially up to 24 months after surgery. CTCs were collected and morphology classified by utilizing a CTC test workflow which uses negative enrichment and immunofluorescence methods to capture and identify CTCs from blood sample. Captured CTCs (epithelial type) were screened with a customized imaging analysis pipeline, a cytological profile of each CTC was created, including cell size, shape, fluorescent intensity and texture etc. Results: The CTC detection rate was 78.1% (78 of 105) prior to surgery, and a total of 726 CTCs were enumerated. Median CTC count number was 3. 5 classes of CTCs with distinct morphological features were observed in lung cancer patients’ CTC tests, briefly, CTC class I and class II possessed large nuclei but relatively lower epithelial expression level, CTC class III, IV, V possessed small nuclei but relatively higher epithelial expression level, CTC class III possessed irregular shaped nuclei, CTC class V possessed relatively lower nuclei/cytoplasm ratio. Class III accounted for the highest proportion of captured CTCs III, about 35.5% with Class I 14.8%, Class II 15.3%,Class IV 17.8% and Class 5 16.6%. Postoperative recurrence and metastasis were observed in 16 patients. CTCs positive were found in 14 patients (87.5%). 145 CTCs were collected, Median CTC count number was 3,Cluster III accounted for 47.3%, with Class I 11.8%,Class II 13.3%,Class IV 14.5% and Class V 11.8%; Patients with Cluster 3 dominant were associated with increased risk of local recurrence ( p < 0.05) and distant metastasis ( p < 0.05). Conclusions: Small and irregular nuclei CTC is significant associated with increased risk of recurrence disease. Morphology Classification of circulating tumor cells is feasible in monitoring the recurrence of disease and may potentially identify the patients who may benefit from further therapy.
Cell-free KRAS in pancreatic cancer as a rapid prognostic marker.
Authors: Saiz-Lopez Patricia , Alcaraz-Ortega Raquel , Lopez Ana Maria , Llorente Beatriz , Diez-Ordoñez Maria Soledad , Pérez Álvarez Juan Carlos , Cáceres Porras Maria José , Palacios Ball Johanna Beatriz , Echevarria-Iturbe Celina , Crespo Guillermo
Published: May 2020

e15534

Background: KRAS mutations are present over 90% of patients with pancreatic cancer (PC). Liquid biopsies provide the opportunity to genotype alterations in a less invasive way and offer a possibility to monitor the molecular characteristics of a cancer through the course of treatment. We aim to establish an association between the mutational levels of KRAS detected in plasma with patient stage and evolution. Methods: Plasma samples were collected from patients with newly diagnosed PC, before any treatment and during one year. Diagnosis of any previous cancer was an exclusion criterion. Samples were frozen (-80ºC) until analysis of KRAS mutations in cell free DNA (cfDNA) by real time PCR. Specifically, Idylla TM technology was used due to the rapid capacity to report up to 21 KRAS mutations in exon 2, 3, and 4 (LOD = < 5%), without necessity of cfDNA isolation. Mutation was considered present (categorical measurement), taking into account cycle quantification values (Cq) established by the commercial test. Cut-off values for Cq were studied by the ROC curve (receiver operating characteristic) with the Youden's J statistic. Paired tissue samples were analyzed when available. Results: A total of 23 patients were enrolled, median age 65 years old, 57% male, 100% adenocarcinoma, 18 (70%) metastatic. Sixteen patients could also be tested in tissue, being 15 (93.8%) positive for KRAS mutation. Plasma basal mutations were detected in 10 patients (43.5%), all in stage IV (p = 0.046), and interestingly, all with liver involvement (p = 0.003). Moreover, these patients also had metastasis in another organ apart from the liver. The most frequent mutation was G12V (5), followed by G12D (4) and G12R (1). Patients with basal KRAS mutations detected in plasma had a survival of 6.1 months (CI 95%, 3.7-8.5), and non-mutated 11.2 months (95% CI, 7.8-14.7) (p = 0.041). The presence of these cell-free KRAS mutations was more informative for survival than stage IV disease vs III (p = 0.054). Paying attention to KRAS wild type detection in plasma samples used as an internal control of the PCR, we could establish a Cq value cut-off of < 21 Cq for mutant samples, with a sensitivity of 100% (95% CI, 75%-100%) and specificity of 50% (95% CI, 19%-81%), area under ROC curve 0.823 (95% CI, 0.649-0.997). Conclusions: Measurement of cell-free KRAS in plasma samples at diagnosis could be used to quickly predict metastasis and survival in patients with pancreatic adenocarcinoma.
The systemic inflammation response index (SIRI) predicts oncological outcome and correlates with tumor burden in metastatic pancreatic cancer.
Authors: Pacheco Barcia Vilma , France Talya , Asselah Jamil , Mondejar Rebeca , Romero-Laorden Nuria , Zogopoulos George , Serrano Rosario , Donnay Olga , Rogado Revuelta Jacobo , Del Campo Lourdes , Martinez Elia , Martin-Perez Elena , Arlanzón Carmen , Guo Katie , Ochoa Pilar , Alcindor Thierry , Barrena Castello Gabriel , Colomer Ramon
Published: May 2020

e15537

Background: The SIRI, defined by neutrophil x monocyte/lymphocyte 109/L, has recently emerged as a prognostic factor for pancreatic cancer. However, the association between SIRI values after chemotherapy and tumor response has not been evaluated. Methods: 161 metastatic pancreatic cancer patients were retrospectively analyzed. Associations between overall survival (OS), chemotherapy and SIRI were analyzed. A larger number of patients with pre-treatment SIRI (pre-SIRI) were collected so, post-treatment SIRI (post-SIRI) evaluated after three cycles of chemotherapy, was adjusted for analysis. Results: Median age 66 years. 59 (36%) received gemcitabine + nab-paclitaxel, 40 (24%) gemcitabine, 22 (17%) mFOLFIRINOX, 13 (7%) other regimens. 27 (16%) had not received treatment. Pre-SIRI≥2.3×109/L was an independent, negative predictor of OS compared to pre-SIRI < 2.3×109/L [5 versus 16 months, Hazard Ratio (HR) 2.87, Confidence Interval (CI) 95% 2.02-4.07, P< 0.0001]. In the whole cohort, we observed SIRI values increased after treatment (median pre-SIRI: 1.6×109/L; post-SIRI: 2.3×109/L; P= 0.007). Thus, we analyzed the association between tumor response measured by RECIST and pre-SIRI and post- SIRI values. Patients with progressive disease (PD) showed a higher pre-SIRI than those who had a response to chemotherapy (2.7×109/L versus 1.2×109/L, respectively; P< 0.001). We also observed a statistically significant increase in post-SIRI values for PD compared to tumor response (3.2×109/L versus 1.7×109/L, respectively; P= 0.012). As observed for pre-SIRI, post-SIRI ≥2.3×109/L showed a shorter OS compared to post-SIRI < 2.3×109/L (8 versus 17 months, respectively; P= 0.016). Furthermore, patients with pre-SIRI ≥2.3×109/L were more likely to benefit from mFOLFIRINOX showing a median OS of 17 months compared to 6 and 4 months for gemcitabine + nab-paclitaxel and gemcitabine, respectively ( P< 0.001). Conversely, there was no difference for pre-SIRI < 2.3×109/L: 15.9 months versus 16.5 and 16, respectively. Conclusions: SIRI≥2.3×109/L was a prognostic factor for metastatic pancreatic cancer. An elevated post-SIRI showed an association with disease progression and a negative impact on survival. Therefore, an increase in SIRI could be related to high tumor burden and be useful to appropriately select patients who would benefit of a more intensive first-line chemotherapy regimen.
A novel method for enrichment and detection of the six major subtypes of EGFR exon 19 deletions simultaneously by capturing mutant-alleles.
Authors: Pu Yue , Lin Rui , Wang Tao
Published: May 2020

e15539

Background: EGFR exon 19 deletions (19DELs) are important precision therapy targets for NSCLC patients. Due to over twenty subtypes of 19DEL mutations, it is usually challenging for detecting different deletion forms in one assay using limited DNA preparation from plasma sample. Methods: Previously we had developed a high sensitivity detection method, namely PEAC, to detect single somatic mutation in one assay. The method used locked¨Cnucleic acid modified short DNA probe to capture mutant fragment under an optimized temperature. In the present study, we have established a multiplexing PEAC approach for simultaneous enrichment and high sensitivity detection of six major subtypes of EGFR 19DELs.The six subtypes overall account for >86% EGFR exon 19 deletions in the patient population. Results: Genomic DNA of EGFR wild-type was mixed with DNA samples of known EGFR 19DEL mutations. The mutation allele fractions of 5%, 1%, and 0.1% for the six 19del subtypes were constructed, which included c.2235-2249del15 p.E746_A750del, c.2236-2250del15 p.E746_A750del, c.2237-2255delinsT p.E746_A752delinsV, c.2235-2249delinsC p.L747_A750delinsP, c.2240-2254del15 p.L747_T751del, and c.2240-2257del18 p.L747_P753delinsS. The mixed DNA samples were sonicated to mimic the fragment distribution of circulating-free DNA (cfDNA) and PCR amplified for exon 19 regions. Multiplexing enrichment using the six specific probes targeted to the 19DEL subtypes were performed to the amplified DNA fragments under the optimal binding conditions. Sanger sequencing all showed detectable variants in the enriched samples starting with 5%, 1%, or 0.1% mutants, which indicated the capacity and high sensitivity of multiplexing PEAC technology. To explore the potential of multiplexing with other EGFR mutants, the 0.1% mixed mutants of 19DEL were PCR amplified for EGFR exon 20 and exon 21 in addition to exon 19, and followed with enrichment and detection of the six 19DEL variants. Sanger sequencing can detect all the six subtypes of 19DELs. Our method is superior to ddPCR in some aspects such as high throughput and low starting DNA amounts. Conclusions: Multiplexing PEAC technology can simultaneously enrich and detect the six major subtypes of EGFR 19 deletions with high sensitivity. It also has potential to multiplex with other EGFR driver mutations such as in exons 18-21. The technology may serve as an attractive method for detecting mutations in liquid biopsies in clinical practice.
Prediction and prognostic validation of colorectal cancer therapeutic effect of WM-S1-030 using pre-clinical CTC animal model.
Authors: Lee Hae Ung , Moon Jai-Hee , Kim Joseph , Kim Seong Keun , Shin Jae Sik , Choi Young Jin , Hong Seung-Woo , Kim Sung Min , Lee Hyun Ho , Park Jae-Chan , Jin Dong-Hoon , Jeon Byung Hee
Published: May 2020

e15557

Background: CTC (Circulating tumor cells) has the advantage of being able to effectively investigate non-invasive diagnosis, prediction, and prognosis for non-surgical carcinoma or tissue sample collection. We have developed WM-S1-030 (WMBIO) as a novel inhibitor for mtRTK (mutant receptor tyrosine kinase) identified important targets for cancer progression and metastasis in various cancers, including colorectal cancer, and have tried to prove the possibility of CTC diagnosis, prediction and prognosis of WM-S1-030 using tumor animal models. Methods: Six of colon cancer cell lines expressing mtRTK (pTyr-mtRTK) were transplanted into Balb.c nude mice and NSG mice to collect peripheral blood when the tumor size reached 1000 mm3. CTCs were quantified in peripheral blood using the Smart Biopsy cell isolator (Cytogen). The expression of pTyr-mtRTK in CTCs was analyzed via the Smart Biopsy Image Analyzer (Cytogen). Colon cancer cell lines expressing pTyr-mtRTK were transplanted into NSG mice and we examined the tumor suppression, the enumeration of CTCs and pTyr-mtRTK expression in CTCs after administration of WM-S1-030. We also identified CTCs in peripheral blood of colon cancer patients and analyzed expression of pTyr-mtRTK in CTCs through Smart Biopsy system. Results: As results of the enumeration of CTCs after tumor transplantation into Balb.c nude mice and NSG mice, the number of CTC was 8 ~ 20 times higher peripheral blood in the NSG model, which indicates innate immune responses are critical for the survival of CTCs. Half of CTCs isolated from the blood expressed pTyr-mtRTK. The tumor size, the number of CTCs, and the expression of pTyr-mtRTK in CTCs were examined after the administration of WM-S1-030. The volume of tumor decreased by 67% in the WM-S1-030 group compared to the vehicle. The number of CTCs was reduced by approximately three times after the treatment of WM-S1-030 and also the number of CTCs expressing pTyr-mtRTK was significantly decreased. CTC detection and pTyr-mtRTK expression analysis in the peripheral blood of colon cancer patients detected 9 CTCs per 10 ml of peripheral blood, of which 11 % of CTCs representing pTyr-mtRTK were confirmed. Conclusions: WM-S1-030 was developed and confirmed in the CTC-based animal model, which implies that the analysis of CTCs is the powerful tool for predicting/ prognosing WM-S1-030 in the pre-clinical analysis. Currently, we are conducting clinical diagnostic validation using CTC in the blood of colorectal cancer patients and preparing for clinical trial of WM-S1-030.
ROS new function for ECT imaging observation of sustained 99mTC-labeled cytarabine in liver cancer patient with hydrogen peroxide.
Authors: Zheng Guoqin , Han Yan , Zhang Jian , Li Bian , Chen Dong , Zhao Ming , Fu Qiang , Wu Yanfei , Yu Baofa ,   Shandong Baofa Cancer Therapeutics, Inc.
Published: May 2020

e15568

Background: To investigate the sustaining time of 99mTc labeled cytarabine in the tumor of liver cancer patients by ROS under ECT imaging instrument. Methods: A liver cancer patient with 2 lesions in the liver was enrolled in this study. After obtaining informed consent and excluding relevant contraindications, two tumors in the liver of the patient were guided by single photon emission computed tomography (ECT). Injecting the drug, the tumor was injected with 99m TC-labeled cytarabine solution as control, the tumor was injected with 99m TC-labeled cytarabine + hydrogen peroxide solution (ROS) as experimental, and the two tumor masses were detected by ECT imaging, with an intensity of 1h as background. The drug retention rate was calculated by caculated of deduction of the baseline (minus background) at 1h, 3h, and 23h. Results: The patient was detected by ECT on the right laid down position. The area of the region of interest was 679 account by Gama detector, and the background level of the liver was 494 account. The acquisition time was about 400 seconds as a standard acquisition. The ECT imaging showed that the labeling rate of 99mTC-labeled cytarabine was about 100%. The drug of 99mTC-labeled cytarabineretentionrates in tumor lesions with ROS at 1 hours, 3 hours, and 23 hours were 97.83%, 81.63%, and 60.72%, respectively; the drug retention rates in tumor lesions without ROS at 1 hours, 3 hours, and 23 hours were 44.74%, 15.87%, and 0, respectively. Conclusions: The ROS can hold drug in the tumor long time after the intratumoral injection of drugs with ROS whichoxidant with the extracellular matrix and interstitial of the tumor to cause degeneration and deformation so that drug of Ara-C sustaining in tumor for a long time, and the tumor cells are also degenerated and necrotic and died, so that high concentrations of cytarabine are embedded between the denatured tumor tissues and cells. The drug slowly released from the inside to the outside to kill tumor cells. IT is further confirmed that the ROS is a sustained release agent intratumorally injection and it can significantly increase the residence time of 99mTc-labeled cytarabine in the tumor, prolong the drug action time of cytarabine, and improve the effect of killing tumor cells.
Effect of hydrogen peroxide on the sustained release of 99mTc-labeled cytarabine in melanoma of C57 mice by ECT imaging pharmacology.
Authors: Han Yan , Fu Qiang , Yu Baofa ,   Shandong Baofa Cancer Therapeutics, Inc.
Published: May 2020

e15569

Background: To investigate the residence time of 99mTc-labeled cytarabine in mouse tumors under ECT imaging. Methods: The subcutaneous solid tumor model of mice was established by injection of melanoma cell suspension into the left forelimb of C57 mice. The tumor-bearing mice were divided into treatment group and control group when the longest diameter of the tumor was 5-8 mm. Group 3 only. The mice in the control group were given intratumoral injection of 0.1 ml of 99mTc-labeled cytarabine solution, and the mice in the treatment group were given intratumoral injection 0.1 ml 99mTc labeled cytarabine combined with hydrogen peroxide solution at 0.25 h, 0.5 h, 1 h. After 2 hours and 19 hours of administration, the radioactivity intensity values were observed by SPECT Apx-409 ECT phenomenon equipment. Results: The T/W values of ECT imaging in 3 mice in the treatment group were higher than those in the control group at 0.25 h, 0.5 h, 1 h, 2 h and 19 h. The T/W values of ECT imaging decreased in the treatment group at different time points. The amplitude was lower, and the T/W values in each period were higher than those in the control group. Conclusions: The sustained release agent hydrogen peroxide can significantly increase the residence time of 99mTc-labeled cytarabine in the tumor. It is speculated that hydrogen peroxide has a positive effect on prolonging the drug action time of cytarabine and improving the effect of killing tumor cells. It showed that H2O2 can hold the Ara-C for a long time in tumor.
IL-6 influences PD-L1 expression in monocytes and macrophages by effecting protein tyrosine phosphatase receptor type O expression in-human breast cancer.
Authors: Chen Lin , Tang Jinhai
Published: May 2020

e15570

Background: We have previously discovered a relationship between the low expression of protein tyrosine phosphatase, receptor type O (PTPRO) in tumor-infiltrating T cells and immunosuppression. The aim of the present study was to investigate the relationship between decreased PTPRO and increased programmed death ligand 1 (PD-L1) in both the peripheral monocytes and tumor-infiltrating macrophages of human breast cancer (BC). Methods: The expression and correlation of all the indices were explored in monocytes and tumor infiltrating macrophages within both human and mice BC. The mechanic regulations were studied by using both in vitro and in vivo studies. Results: We found a significant decrease in PTPRO in BC peripheral monocytes that was associated with increased PD-L1 expression in peripheral monocytes and tumor-associated macrophages (TAMs) in BC. Monocyte PD-L1 and PTPRO therefore could serve as valuable prognostic indicators for post-surgery patients with BC and were associated with increased T cell exhaustion (Tim3+ T cells). A depletion of PTPRO promoted PD-L1 secretion in both monocytes and macrophages through the JAK2/STAT1 and JAK2/STAT3/c-MYC pathways. Increased IL-6 expression was associated with activation of JAK2/STAT3/c-MYC and with decreased PTPRO expression through the STAT3/c-MYC/miR-25-3p axis. Monocytes and TAMs showed significantly increased miR-25-3p expression, which could target the 3' untranslated region of PTPRO. The miR-25-3p expression positively correlated with serum IL-6 levels, but inversely correlated with PTPRO in BC monocytes. IL-6/STAT3/c-MYC activation enhanced in vitro miR-25-3p transcription and decreased PTPRO, while further promoting PD-L1 secretion. Adoptive cell transfer of c-MYC/miR-25-3p–modified monocytes promoted tumor growth by downregulating PTPRO and causing a PD-L1–induced immunosuppression in breast tumor model. Conclusions: Increased serum IL-6 downregulated PTPRO expression in BC monocytes and macrophages by activating STAT3/c-MYC/miR-25-3p and by further enhancing PD-L1 expression through JAK2/STAT1 and JAK2/STAT3/c-MYC signaling.
Use of real-world clinical lab data to reveal Asian NTRK fusion test availability patterns.
Authors: Eckstein Markus , Bloom Kenneth Joel , Fosvig Susanne Munksted , Hoefsmit Marieke , Clark Jordan
Published: May 2020

e15574

Background: The development and launch of first-generation tropomyosin receptor kinase (TRK) inhibitors, such as larotrectinib and entrectinib, has brought targeted treatment options to neurotrophic tropomyosin-related kinase (NTRK) fusion–positive adult and pediatric cancer patients who may have been out of treatment options. Widespread adoption of NTRK fusion testing is needed to support successful selection of patients for these TRK inhibitors as well as other targeted therapies. The objective of this study was to investigate global availability of NTRK fusion testing, with a specific focus on Asia. Methods: For this study, real-world clinical pathology laboratory data, derived from Diaceutics’ proprietary Global Diagnostic Index (GDI) of more than 2500 labs, were investigated to assess the current footprint of NTRK fusion testing in Asia. Availability of clinical routine testing was analyzed in selected cohorts of laboratories across different regions, comparing Asia to the US and Europe. Laboratories were included based on their large volume of testing or on their importance (e.g., large academic and large commercial laboratories). Results: Data on availability of NTRK fusion testing by clinical laboratories reveal different patterns of NTRK fusion test adoption in Asia, depending on the region under study. In China and similarly in the US, Italy, and France clinical NTRK fusion testing is already being performed by the majority of laboratories from the selected cohort. In Japan, physicians have limited access to testing, with only a few laboratories from the selected cohort performing testing for NTRK fusions. Conclusions: NTRK fusion testing is essential for identifying patients eligible for first-generation TRK-inhibitors. Various laboratories across selected cohorts of labs in China and Japan lack the capability to initiate clinical NTRK fusion testing in-house, denying some patients the potential benefit from these breakthrough therapies. Consequently, patient outcomes will be suboptimal due to lack of testing infrastructure. The readiness of Asian markets to support patient selection successfully can affect the rate of testing and the impact of these innovative therapies on patients’ lives. [Table: see text]
Baseline radiomics features (RF) in metastatic colorectal cancer (mCRC): Correlation with m site and clinical-pathological characteristics.
Authors: Vivolo Raffaella , Boldrini Luca , Lenkowicz Jacopo , Caliolo Gianluca , Camarda Floriana , Anghelone Annunziato , Di Stefano Brunella , Calegari Maria Alessandra , Pozzo Carmelo , Basso Michele , Liguori Carolina , De Gaetano Anna Maria , Dinapoli Nicola , Manfredi Riccardo , Valentini Vincenzo , Tortora Giampaolo , Salvatore Lisa
Published: May 2020

e15589

Background: R is an emerging field of research based on the extraction of a large amount of F from biomedical images and on computed analysis algorithms of tumour architecture. Few data regarding mCRC are available, in particular no correlation of baseline RF both with m sites and clinical-pathological characteristics was so far investigated. Methods: Baseline chest-abdomen CT scans of mCRC patients (pts) were retrospectively analysed. RF were extracted from Regions of Interest (ROI) delineated on CT scan from each m sites, including primary tumor, when on site. The association of specific F and disease site (liver, lung, nodes, peritoneum and on-site primary tumor) was investigated. Sites similarity was assessed with Principal Component Analysis, an unsupervised learning technique to identify patterns and clusters. Then RFs were tested individually for correlation with clinical-pathological covariates of interest (gender, CEA level, synchronous disease, RAS/BRAF status, mucinous histology, grading, number of m site, primary tumor site). Wilcoxon-Mann-Whitney test was used for this purpose (significance level set at 0.05). Results: After RF extraction from the different ROIs, the dataset was composed of 433 observations of 236 variables. Observations referred to the number (N) of pts = 89 and the N of ROIs = 18. RF classes were divided in statistical F (grey-level histogram) (N of F = 10); morphological F (N = 14); texture F GLCM (grey level co-occurrence matrix) (N = 100); texture F GLRLM (grey level run length matrix) (N = 66); texture F GLSZM (grey level size zone matrix) (N = 32). Regarding the association of RF with m sites, an homogenous distribution with liver, nodes, peritoneum and primary tumor was detected, while lung metastases showed a different pattern for all the RF classes. A significant correlation of specific RF with clinical-pathologic characteristics was shown, in particular with gender, CEA level, synchronous disease, mucinous histology, RAS/BRAF status. Conclusions: Despite its retrospective nature and the limited number of pts, this is the first experience demonstrating I) a different pattern of RF for lung m versus an homogeneous RF distribution for the other m sites; and II) a significant association of specific RF with few clinical-pathologic characteristics. Our results, if confirmed in a prospective validation set, may represent an hypothesis generator regarding the different behaviour of lung metastases and a possible R signature able to identify different prognostic subgroups of pts.
Effect and mechanism of ikkβ on proliferation, apoptosis, and metastasis of breast cancer cells with different expression levels of leptin and its receptors.
Authors: Kong Yan , Dong Qian , Geng Cuizhi , Jiang Da
Published: May 2020

e15592

Background: To investigate the effect and mechanism of IKKβ on proliferation, apoptosis, invasion and metastasis of breast cancer cells with different expression levels of leptin and ObR. Methods: IKKβ in breast cancer cells was knocked down via virus transfection technology. MTT, flow cytometry, cell scratch test and clone formation test were used to observe the effect of IKKβ on proliferation, apoptosis and metastasis of breast cancer cells with different expression levels of leptin and ObR. RT-PCR was used to detect the downstream genes expressions in IKKβ signaling pathway. Results: The expression levels of leptin and ObR in MDA-MB-231 cells were higher than those in MCF-7 cells. Lentivirus can successfully infect breast cancer cells in vitro and knock down the expression of IKKβ. Knocking down IKKβ reduced the viability of MDA-MB-231 cells, but had little effect on MCF-7, and had an effect on the cell cycle of both cells. After knocking down IKKβ, the apoptotic rate of MDA-MB-231 cells increased. There was no significant difference in the apoptotic rate of MCF-7 cells. The number of clones of MCF-7 and MDA-MB-231 cells decreased, and the difference was significant. The effect on MDA-MB-231 cells was more significant. The scratch mobility of MDA-MB-231 cells decreased significantly, with significant difference.After knocking down IKKβ, the expressions of leptin and ObR, IKBKG, P65 and HIF in MCF-7 cells were significantly decreased, and there was statistical significance. However, the above indicators were significantly increased in MDA-MB-231 cells. The expression level of IKBKG was increased significantly, whereas that of HIF did not differ significantly, as compared with that of control group. Conclusions: The proliferation, metastasis and apoptosis of MDA-MB-231 cells were inhibited after blocking the IKKβ pathway, while the proliferation of MCF-7 cells was inhibited, and the apoptosis did not change significantly. The difference between the two cells may be related to the different expression levels of leptin and ObR.After blocking the IKKβ signaling pathway, the expressions of genes related to the IKKβ signaling pathway in MDA-MB-231 cells were increased, whereas those in MCF-7 cells were decreased, which might be related to the different expression levels of leptin and ObR.After knocking down IKKβ, the expressions of HIF gene in MCF-7 cells were decreased, whereas those in MDA-MB-231 cell lines were not affected. It needs to be verified whether this difference is related to leptin and ObR.
Chick chorioallantoic membrane (CAM) assays as a model of xenografts derived from circulating cancer stem cells from breast cancer patients.
Authors: Pizon Monika , Schott Dorothea , Pachmann Ulrich A. , Pachmann Katharina , Schobert Rainer
Published: May 2020

e15596

Background: Circulating cancer stem cells (cCSCs) are a rare fraction of circulating tumor cells with a profile of stemness, resistance to chemotherapy and the capacity to generate metastases. Patient-derived xenografts (PDX) are an increasingly noticed tool in oncology, providing biologically meaningful models of many cancer types, and potential platforms for the development of precision oncology approaches. Commonly, mouse models are used for the in vivo assessment of potential new therapeutic targets in cancers. However, given the high cost, time investment, and legal obligations of such animal models CAM assays are an attractive alternative. Methods: In this study, primary cultures from circulating cancer stem cells were established using sphere-forming assays. Subsequently, tumorspheres were transplanted onto the CAM membrane of fertilized chicken eggs to form secondary microtumors. Results: We have developed an innovative in vitro platform for cultivation of CSCs from peripheral blood of breast cancer patients. The number of tumorspheres increased significantly with tumor progression. Patients with metastatic disease had statistically more tumorspheres as compared to patients without metastasis (30 vs 10/100µl blood, p < 0.05). Patients with multiple metastases had more tumorspheres compared to patients with single metastases (30 vs 60/100µl blood, p < 0.05). The number of tumorspheres was positively correlated with Ki-67, Her2 status and grade score in primary breast tumors. Tumorspheres showed self-renewal, growth potentials, invasion and differentiation in vivo. Their transplantation on CAM membranes resulted in the rapid formation of microtumors in many cases. These tumors pathologically closely resembled the primary tumor. The success rate of PDX was positively correlated with aggressiveness and proliferation capacity of the primary tumor. Conclusions: The number of tumorspheres cultured from peripheral blood of cancer patients and the success rate of establishing PDX directly reflect the aggressiveness and proliferation capacity of the primary tumor. A CAM-based PDX model using circulating cancer stem cells provides a fast, low-cost, easy to handle and powerful preclinical platform for drug screening, therapy optimization, and biomarker discovery.