Background

Due to the adverse side effects of current cancer chemotherapeutics development of targeted anti-cancer medications is under intensive focus. In the present study a fusion protein consisting of the catalytic and translocation domains of diphtheria toxin fused to BR2, a cancer specific cell penetrating peptide, was produced as a new targeted anti-cancer agent and its cytotoxic effects on various cell lines including MCF-7 and HeLa (as cancerous), and HEK293 and HUVEC (as normal) cell lines was evaluated.

Methods

DT386-BR2 protein was expressed in E. coli cells followed by protein purification using affinity chromatography and authenticated by SDS-PAGE and Western blotting. Next, the purified protein was subjected to evaluation of its cyto-lethal effects on the mentioned cell lines at various concentrations. DT386 and BR2 alone were also produced and used as negative controls. In this regard, MTT assay followed by flow cytometry were used for cytotoxicity assessment and prediction of induced cell death mechanism.

Results

SDS-PAGE and Western blotting confirmed expression of DT386-BR2 fusion protein by revealing a band of about 47kDa. In case of the cancerous cell lines, i.e. HeLa and MCF-7 cells, significant reduction in survival percent was shown when compared to the controls. However, no significant cytotoxicity was observed in case of the normal cell lines, HUVEC and HEK293 cells. In addition, DT386 and BR2 alone did not show any significant cytolethal effects. The IC50 of DT386-BR2 for HeLa and MCF-7 was about 0.55 and 2.08 μg/ml, respectively. Furthermore, flow cytometry revealed dose dependent apoptosis induction by DT386-BR2 after 12 hrs.

Conclusions

This chimeric protein showed promising cancer cell specific cytotoxicity and might be considered as a new drug candidate for treatment of some types of cancer. Further studies regarding non-specific cytotoxicity and in vivo animal and preclinical studies of this protein are undergoing.

Legal entity responsible for the study

Isfahan University of Medical Sciences

Funding

Research Deputy of Isfahan University of Medical Sciences

Disclosure

All authors have declared no conflicts of interest.

Background

Elderly cancer patients aged 65 or more are generally frail compared with younger patients. They are considered to be the age group of less fitting for clinical trials, especially investigating feasibility. For the past decade, the types of the agents have been changing and the number of drug classes has been increasing. The aim of our study was to investigate the impact by age on dose-limiting toxicities (DLT) in each type of agents.

Methods

Retrospective analysis was implemented on patients who participated in phase 1 oncology trials between 1995 and 2016 in our institution. DLT was defined in each trial protocol. The rate of DLT was compared between younger patients (< 65 year) and elderly patients (≥ 65 year) for cytotoxic agents and molecular targeted agents.

Results

In this period, 973 patients underwent 214 enrolments in trials of cytotoxic agents and 996 enrolments for molecular targeted agents. For cytotoxic agents, 165 enrolments with 23 DLT events (13.9%) occurred in the younger group and 49 enrolments with 13 DLTs (26.5%) in the elderly group (p = 0.05). With molecular targeted agents, 728 enrolments with 57 DLTs (7.8%) occurred in the younger group and 268 enrolments with 31 DLTs (11.6%) in elderly group (p = 0.08).

Conclusions

In phase 1 trials, enrolment of elderly patients should be carefully discussed especially in trials of cytotoxic agents. With molecular targeted agents, age groups exert less impact on DLT.

Legal entity responsible for the study

National Cancer Center Hospital

Funding

N/A

Disclosure

S. Iwasa: Grants and personal fees from Eli Lily, personal fees from Taiho, grants and personal fees from Chugai, grants from Novartis, grants from Merck-Serono, grants from Daiichi-Sankyo, grants from Bristol-Myers Squibb, from null, outside the submitted work. Y. Fujiwara: Grants from AbbVie, grants and personal fees from AstraZeneca, grants from Chugai, grants from Daiichi-Sankyo, grants from Eisai, grants from Eli Lilly, grants from Incyte, grants from Merck Serono, grants and personal fees from MSD, grants from Novartis, personal fees from Taiho, grants and personal fees from BMS, personal fees from Ono, outside the submitted work. T. Shimizu: Research expenses from Bristol-Myers Squibb, Daiichi-Sankyo, PharmaMar, FivePrime, 3D Medicine, Takeda-Milleniam, personal fees (lecture fees) from Chugai Pharmaceutical CO., LTD., Taiho Pharmaceutical Co., Ltd., Ono Pharmaceutical Co., Ltd., Ono Pharma Taiwan Co., Ltd., Boehringer Ingelheim, outside the submitted work. N. Yamamoto: Grant from Bristol-Myers Squibb, Chugai, Taiho, Eisai, Lilly, Quintiles, Astellas, Novartis, Daiichi-Sankyo, Pfizer, Boehringer ingelheim, Kyowahakko Kirin, Bayer, Ono Pharmaceutical Co, Ltd and Takeda, other from Chugai., Ono Pharmaceutical Co., Ltd., AstraZeneca, Pfizer, Lilly and Bristol-Myers Squibb outside the submitted work. All other authors have declared no conflicts of interest.

Background

Lymph nodes (LNs) are an essential organ in an immune system, and lymphatic vessels connect them. Their primary function is to maintain the health of the body via filtering morbific material from the lymphatic fluid. In the case of cancer metastasis, cancer cells are disseminated from primary tumor via the bloodstream to the distant sites. Clinically, detection of metastasis within sentinel lymph node (SLNs) is significantly predictive of prognosis with implications for the patient's survival and adjuvant therapy. The limited studies available report that dissection of normal lymph nodes (LNs) is involved in the activation and rapid growth of latent tumors in distant metastasis.

Methods

Here we show that dissection of LNs with and without tumor cells may activate distant metastasis in the mice. Tumor cells were inoculated intravenously and intranodally into MXH10/Mp/lpr-lpr mice to create a metastatic lung mouse model.

Results

Bioluminescence images indicated tumor cells in the lung were activated after dissection of subiliac lymph node (SiLN) with and without tumor cells. While, no luciferase activities were detected in the lung without dissection group excluding intravenous inoculation group.

Conclusions

The results indicated that LN dissection might affect secondary tumor formation, and the metastatic lymph node dissection may increase metastasis if the nodes might contain tumor cells. Furthermore, removal of normal lymph nodes may induce metastases in the distant organs if the organs contain tumor cells. Our results indicate dissection/biopsy of lymph node metastasis may have the potential hazard of inducing metastases in the distant organs.

Legal entity responsible for the study

N/A

Disclosure

All authors have declared no conflicts of interest.

Background

Obesity and High body mass index are associated with a higher risk of developing renal cell carcinoma (RCC). Leptin is a peptide hormone produced predominantly by adipocytes that is elevated in obese individuals. The main function of leptin is to regulate body weight and appetite. Leptin may play a role in carcinogenesis of many cancers including RCC. We aimed to investigate the role of leptin Receptor gene (A/G) polymorphism (rs1137101) in RCC.

Methods

This study was carried out by cooperation between Clinical Oncology & Nuclear Medicine, Medical Biochemistry and Internal Medicine Departments, Faculty of Medicine, Menoufia University in the period from May 2015 to April 2017. It was conducted on 123 individuals classified into; group I: included 73 patients with histological diagnosis of RCC and group II: 50 healthy control subjects. For RCC patients Staging was done according to the American Joint Committee on Cancer: the 7th edition, stage IV patients received SUTENT® (sunitinib malate) as first line treatment. Genotyping of the Gln223Arg (rs1137101) A/G polymorphism was measured by Real-Time PCR and compared between both groups and correlated with different patients, disease characteristics and survival.

Results

This study included 66 males and 57 females, the mean age 58.65 + 8.62. The results of this study revealed that there was a significant statistical difference as regards genotype frequency of LEPR gene A/G polymorphism between the two studied groups, with the highest frequency of GG genotype among RCC patients (67.1%), while AA genotype had the highest frequency in the control group (60%); (p < 0.001). In RCC patients with GG genotype is associated with more advanced stage (stage III and IV) (46.9%) compared to GA & AA genotypes (12.5%), and higher nuclear grade G3 (28.5%) compared to GA, AA genotypes (4.2%). GG genotype has worse survival versus GA and AA genotypes; p value <0.001.

Conclusions

Leptin Receptor gene (A/G) polymorphism (rs1137101) might be a candidate risk factor for developing RCC. In RCC patients with GG genotype is associated with more advanced stage and higher nuclear grade and shorter survival compared with patients with GA & AA genotypes.

Legal entity responsible for the study

Menoufia University

Funding

Has not received any funding

Disclosure

All authors have declared no conflicts of interest.

Background

Radiotherapy is one of the commonly used approaches to treat cancer. The research trend and breakthroughs in radiotherapy is focusing on the high linear energy transfer (LET) radiation like the alpha particles. Thus, the alpha particles based method is an important option of radiotherapy development, and Targeted Alpha Therapy (TAT) is the most important application of using alpha particles in the past decade. This paper highlights the research work done on computational modelling of TAT at Canadian Nuclear Laboratories (CNL).

Methods

Mathematical model is developed to study the blood flow through a two-dimensional vasculature embedded in a solid tumour or a normal cell. A mesoscale modeling (with a length scale of 1 µm – 1mm and a time scale of 1 ns - 1 µs) is conducted in order to gain an understanding of dynamic alpha-immuno-conjugate (AIC)-attached blood flow interaction in vasculature and its influence on AIC motion in the main flow direction and the perturbations/instabilities of the AIC trajectories in the transverse direction. Microdosimetic modeling of TAT using Monte Carlo toolkit to calculate the amounts of the energy deposited in a simplified DNA model of cancer cells and evaluate the absorbed background dose for normal cells. Coupled the computational fluid dynamic (CFD) simulation with the Monte Carlo prediction to evaluate the efficacy of the AIC during transport process of killing tumour cells and the toxicity to the surrounding normal cells. Post-processing large amount of data calculated by high fidelity CFD simulation and microdosimetric analysis and help review the efficacy of the TAT treatment that is invisible in experiment in pre-clinical stage using a powerful visualization tool.

Results

A coupled model based on the Geant4 Monte Carlo micro-dosimetry technique and Computational Fluid Dynamics analysis was established. The transient drug delivery process and background dose to the cells along the pathway were investigated. A mesoscale numerical simulation in a simple 2D capillary was performed to determine the transient toxicity of the Alpha-Immuno-Conjugate to the DNA of a targeted cell.

Conclusions

The paper demonstrates the feasibility of coupling CFD simulations and microdosimetic modeling to evaluate the efficacy of the TAT realistically and accurately.

Legal entity responsible for the study

Canadian Nuclear Laboratories (CNL)

Funding

Has not received any funding

Disclosure

The author has declared no conflicts of interest.

Background

Patient Preference Assessment Tool (PPAT) is a patient self-assessment questionnaire designed by Emory (US) to allow easy and rapid evaluation of patient preferences for phase 1 (P1) trial participation. Beyond being a legal requirement, informed consent has a critical role to ensure that patient’s trial participation trial matches his/her expectations and understanding of treatment options. This tool has not been independently validated yet. We therefore aimed at assessing PPAT in two cohorts of P1 and phase 2-3 (P2-3) patients (pts) treated at Gustave Roussy (France).

Methods

PPAT was translated in French and proposed to: (1) a cohort of P1 pts just before their first clinic in the P1 department, i.e. prior to trial discussion and inclusion; and (2) an independent cohort of pts recently (< 1 month) included in a P2-3 trial for advanced disease. PPAT was distributed by nurses; pts had to answer alone, by choosing 4 out of 17 proposed items, including one free commentary box.

Results

Between December 2015 and September 2017, 51 P1 pts and 28 P2-3 pts with various tumor types agreed to fill in PPAT; 92% of them adequately completed the questionnaire. Answers to PPAT were similar between both cohorts, with no difference in any item reaching statistical significance (Chi-2 test). The three most frequent answers of our pts were: “Fighting my cancer as much as possible”, “Accessing the best treatment”, “Getting the best possible care from expert doctors” with respectively 78.4%, 64.7% and 47.1% in the P1 cohort and 79%, 61% and 64% for P2-3 pts. By contrast, the three most frequent answers in Emory’s cohort were “Feeling as good as possible every day given my condition”, “Fighting my cancer as much as possible” and “Avoiding spending lots of hours in the clinic”. Five pts answered the “free” item.

Conclusions

Phase I patient preferences are different between Emory and Gustave Roussy’s pts, but P1 pts' answers do not differ from P2-3 pts at Gustave Roussy. PPAT is a well-suited patient-friendly questionnaire, whose utility could be further assessed in populations from alternative cultural or ethnic backgrounds.

Legal entity responsible for the study

Gustave Roussy

Funding

Gustave Roussy

Disclosure

All authors have declared no conflicts of interest.

Background

Legal entity responsible for the study

N/A

Funding

Has not received any funding

Disclosure

The author has declared no conflicts of interest.