BackgroundMetastasis is a process by which cancer cells learn to form satellite tumors in distant organs and represents the principle cause of death of patients with solid tumors. NSCLC is the most lethal human cancer due to its high rate of metastasis.Methodology/Principal FindingsLack of a suitable animal model has so far hampered analysis of metastatic progression. We have examined c-MYC for its ability to induce metastasis in a C-RAF-driven mouse model for non-small-cell lung cancer. c-MYC alone induced frank tumor growth only after long latency at which time secondary mutations in K-Ras or LKB1 were detected reminiscent of human NSCLC. Combination with C-RAF led to immediate acceleration of tumor growth, conversion to papillary epithelial cells and angiogenic switch induction. Moreover, addition of c-MYC was sufficient to induce macrometastasis in liver and lymph nodes with short latency associated with lineage switch events. Thus we have generated the first conditional model for metastasis of NSCLC and identified a gene, c-MYC that is able to orchestrate all steps of this process.Conclusions/SignificancePotential markers for detection of metastasis were identified and validated for diagnosis of human biopsies. These markers may represent targets for future therapeutic intervention as they include genes such as Gata4 that are exclusively expressed during lung development.

ABSTRACTment of MM cells leads to significant cytotoxic effects through induction of apoptosis as the main mode of action and activation of complement-dependent cytotoxicity as a second killing mechanism. Furthermore, GRP78 is an interesting target in MM due to its sensor function in the unfolded protein response pathway. 13,18 PAT-SM6 has been further investigated in various cancer animal models and has been shown to induce significant reductions in metastasis and/or tumor size and volume. 19,20 In formal toxicology evaluations, PAT-SM6 showed no adverse effects up to repeated doses of 50 mg/kg in cynomolgus monkey studies. PAT-SM6 appears to be safe and well tolerated and demonstrated encouraging preliminary anti-tumor activity when used at low doses in patients with recurrent 'in-transit' cutaneous melanoma. 20Based on these promising preclinical observations, the clinical study of PAT-SM6 in MM was initiated. Methods PatientsAdult patients with relapsed and/or refractory MM who had received at least two previous lines of treatment including an immunomodulatory agent (thalidomide or lenalidomide) and a proteasome inhibitor and were showing progressive disease, as defined by the International Myeloma Working Group (IMWG) criteria 21 were included. Patients had Eastern Cooperative Oncology Group (ECOG) performance status grade 0 -2 at study entry, an absolute neutrophil count ≥1000/mm 3 , a platelet count ≥30x10 9 /L, hemoglobin level >8 g/dL and adequate renal function, with a calculated creatinine clearance (using the Cockcroft-Gault formula) of >30 mL/min. Study design and treatmentThis study was approved by the Paul-Ehrlich Institute and the local ethics committee of the University of Würzburg and conducted in accordance with the Declaration of Helsinki.The primary study objective was to assess the safety and tolerability of escalating doses of the anti-GRP78 monoclonal antibody PAT-SM6 in subjects with relapsed or refractory MM. The secondary objectives were to evaluate the efficacy and pharmacodynamics, exploratory biomarkers and anti-PAT-SM6 antibodies. The study also aimed to evaluate the duration of response and progression-free survival.PAT-SM6 was administered intravenously over 90 min. Patients in four dose cohorts (0.3, 1, 3 and 6 mg/kg) were treated on days 1, 3, 8 and 10 (Figure 1). Due to limited amount of antibody product available, the study was designed as a small pilot trial of four increasing doses rather than an open trial with ongoing dose escalation and treatment of patients until progression. Patients received pre-medication with antihistaminic drugs. Prior to each dose escalation, the DSMB met to review the tolerability and toxicity in the previous dose group.A screening examination was performed within 14 days prior to dosing and subjects returned for outpatient evaluations on days 15, 22, 29 and 36 for safety and response and for pharmacodynamic assessments. Criteria for evaluationSafety parameters were evaluated using Common Terminology Criteria for Adverse Events (CTCAE) criteria. ...

Human lung cancer is a disease with high incidence and accounts for most cancer-related deaths in both men and women. Metastasis is a common event in non-small cell lung carcinoma (NSCLC), diminishing the survival chance of the patients with this type of tumor. It has been shown that MYC is involved in the development of metastasis from NSCLC, but the mechanism underlying this switch remained to be identified. Here, we focus on GATA4 as a MYC target in the development of metastasis with origin in lung adenocarcinoma, the most common type of NSCLC. Epigenetic alterations at the GATA4 promoter level were observed after MYC expression in lung adenocarcinoma in vivo and in vitro. Such alterations include site-specific demethylation that accompanies the displacement of the MYC-associated zinc finger protein (MAZ) from the GATA4 promoter, which leads to GATA4 expression. Histone modification analysis of the GATA4 promoter revealed a switch from repressive histone marks to active histone marks after MYC binding, which corresponds to active GATA4 expression. Our results thus identify a novel epigenetic mechanism by which MYC activates GATA4 leading to metastasis in lung adenocarcinoma, suggesting novel potential targets for the development of antimetastatic therapy.

<p>PDF file - 227K, GATA4 promoter analysis and in-vivo material used for next generation-sequencing</p>

<p>PDF file - 68K, Statistical significance of methylation percentage differences at GATA4 promoter of GATA4 positive and negative lung tumors and GATA4 positive liver metastasis tissues.</p>