Prognostic significance of germline BRCA mutations in patients with HER2-POSITIVE breast cancer.

Background: HER2-positive breast cancers are rare amongst BRCA mutation carriers. No data exist regarding clinicopathological characteristics and prognosis of this subgroup of patients. Materials and methods: Using a retrospective matched cohort design, we collected data from 700 women who were diagnosed with operable invasive breast cancer from January 2006 to December 2016 and were screened for germline BRCA mutations. Clinicopathological features and survival rates were analyzed by BRCA and HER2 status. Results: One hundred and fifteen HER2-positive/BRCA mutated cases were evaluated in comparison to the three control groups: HER2-positive/BRCA wild type (n = 129), HER2-negative/BRCA mutated (n = 222), HER2-negative/BRCA wild type (n = 234). HER2-positive breast cancers were more likely to have high histologic grade and high proliferation rate than HER2-negative neoplasms, regardless of BRCA mutation status. An interaction between BRCA mutations and HER2-positive status was found to correlate with worse survival after adjusting for prognostic variables (HR = 3.4; 95% CI: 1.3-16.7). Conclusions: Co-occurrence of BRCA mutations and HER2-positive status is a poor prognostic factor in patients with early or locally advanced breast cancer. This finding may be a proof of concept that a combined pharmacological intervention directed to these targets could be synergistic

EGFR genomic alterations in cancer: prognostic and predictive values

none11: The role of EGFR in cancer development and progression has been recognized for long time in a variety of human malignancies including lung, head and neck, colon, breast, ovary and glioma. Recently its role as a target of antineoplastic agents has also been identified and a variety of EGFR-targeted drugs is already being used in a clinical setting and others are at present under investigation. Many data involving EGFR protein expression are now available for the choice of anti-EGFR monoclonal antibodies in colorectal cancer and with regard to EGFR gene mutations for the choice of tyrosine kinase inhibitors in lung cancer. Other EGFR-related molecular factors, including the EGFR gene copy number, are currently under investigation. This review summarizes both preclinical and clinical available data regarding EGFR genomic alterations as prognostic and predictive factors.openBronte, Giuseppe; Terrasi, Marianna; Rizzo, Sergio; Sivestris, Nicola; Ficorella, Corrado; Cajozzo, Massimo; Di Gaudio, Francesca; Gulotta, Gaspare; Siragusa, Sergio; Gebbia, Nicola; Russo, AntonioBronte, Giuseppe; Terrasi, Marianna; Rizzo, Sergio; Sivestris, Nicola; Ficorella, Corrado; Cajozzo, Massimo; Di Gaudio, Francesca; Gulotta, Gaspare; Siragusa, Sergio; Gebbia, Nicola; Russo, Antoni

Molecular Mechanisms of Trastuzumab Resistance in HER2 Overexpressing Breast Cancer

The epidermal growth factor receptor 2 (HER2) is a tyrosine kinase overexpressed in nearly 20% to 25% of invasive breast cancers. Trastuzumab is a humanized monoclonal antibody that targets HER2. The majority of patients with metastatic breast cancer initially respond to trastuzumab, however, within 1 year of treatment disease progresses. Several molecular mechanisms have been described as contributing to the development of trastuzumab resistance. They could be grouped as impaired access of trastuzumab to HER2, upregulation of HER2 downstream signaling pathways, signaling of alternative pathways, and impaired immune antitumor mechanisms. However, since many of them have overlapping effects, it would be of great clinical impact to identify the principal signaling pathways involved in drug resistance. Significant efforts are being applied to find other therapeutic modalities besides trastuzumab treatment to be used alone or in combination with current modalities

Clinical Considerations of BRCA1- and BRCA2-Mutation Carriers: A Review

Individuals who carry an inherited mutation in the breast cancer 1 (BRCA1) and BRCA2 genes have a significant risk of developing breast and ovarian cancer over the course of their lifetime. As a result, there are important considerations for the clinician in the counseling, followup and management of mutation carriers. This review outlines salient aspects in the approach to patients at high risk of developing breast and ovarian cancer, including criteria for genetic testing, screening guidelines, surgical prophylaxis, and chemoprevention

Combining PARP Inhibition with Platinum, Ruthenium or Gold Complexes for Cancer Therapy

Platinum drugs are heavily used first-line chemotherapeutic agents for many solid tumours and have stimulated substantial interest in the biological activity of DNA-binding metal complexes. These complexes generate DNA lesions which trigger the activation of DNA damage response (DDR) pathways that are essential to maintain genomic integrity. Cancer cells exploit this intrinsic DNA repair network to counteract many types of chemotherapies. Now, advances in the molecular biology of cancer has paved the way for the combination of DDR inhibitors such as poly (ADP-ribose) polymerase (PARP) inhibitors (PARPi) and agents that induce high levels of DNA replication stress or single-strand break damage for synergistic cancer cell killing. In this review, we summarise early-stage, preclinical and clinical findings exploring platinum and emerging ruthenium anti-cancer complexes alongside PARPi in combination therapy for cancer and also describe emerging work on the ability of ruthenium and gold complexes to directly inhibit PARP activity

Impairing energy metabolism in solid tumors through agents targeting oncogenic signaling pathways

Cell metabolic reprogramming is one of the main hallmarks of cancer and many oncogenic pathways that drive the cancer-promoting signals also drive the altered metabolism. This review focuses on recent data on the use of oncogene-targeting agents as potential modulators of deregulated metabolism in different solid cancers. Many drugs, originally designed to inhibit a specific target, then have turned out to have different effects involving also cell metabolism, which may contribute to the mechanisms underlying the growth inhibitory activity of these drugs. Metabolic reprogramming may also represent a way by which cancer cells escape from the selective pressure of targeted drugs and become resistant. Here we discuss how targeting metabolism could emerge as a new effective strategy to overcome such resistance. Finally, accumulating evidence indicates that cancer metabolic rewiring may have profound effects on tumor-infiltrating immune cells. Modulating cancer metabolic pathways through oncogene-targeting agents may not only restore more favorable conditions for proper lymphocytes activation, but also increase the persistence of memory T cells, thereby improving the efficacy of immune-surveillance

Preliminary insights on the mutational spectrum of BRCA1 and BRCA2 genes in Pakhtun ethnicity breast cancer patients from Khyber Pakhtunkhwa (KP), Pakistan

Gene mutations are a source of genetic instability which fuels the progression of cancer. Mutations in BRCA1 and BRCA2 are considered as major drivers in the progression of breast cancer and their detection indispensable for devising therapeutic and management approaches. The current study aims to identify novel pathogenic and recurrent mutations in BRCA1 and BRCA2 in Pakhtun population from the Khyber Pakhtunkhwa. To determine the BRCA1 and BRCA2 pathogenic mutation prevalence in Pakhtun population from KP, whole exome sequencing of 19 patients along with 6 normal FFPE embedded blocks were performed. The pathogenicity of the mutations were determined and they were further correlated with different hormonal, sociogenetic and clinicopathological features. We obtained a total of 10 mutations (5 somatic and 5 germline) in BRCA1 while 27 mutations (24 somatic and 3 germline) for BRCA2. Five and seventeen pathogenic or deleterious mutations were identified in BRCA1 and BRCA2 respectively by examining the mutational spectrum through SIFT, PolyPhen-2 and Mutation Taster. Among the SNVs, BRCA1 p.P824L, BRCA2 p. P153Q, p.I180F, p.D559Y, p.G1529R, p.L1576F, p.E2229K were identified as mutations of the interaction sites as predicted by the deep algorithm based ISPRED-SEQ prediction tool. SAAFEQ-SEQ web-based algorithm was used to calculate the changes in free energy and effect of SNVs on protein stability. All SNVs were found to have a destabilizing effect on the protein. ConSurf database was used to determine the evolutionary conservation scores and nature of the mutated residues. Gromacs 4.5 was used for the molecular simulations. Ramachandran plots were generated using procheck server. STRING and GeneMania was used for prediction of the gene interactions. The highest number of mutations (BRCA1 7/10, 70 %) were on exon 9 and (BRCA2, 11/27; 40 %) were on exon 11. 40 % and 60 % of the BRCA2 mutations were associated Grade 2 and Grade 3 tumors respectively. The present study reveals unique BRCA1 and BRCA2 mutations in Pakhtun population. We further suggest sequencing of the large cohorts for further characterizing the pathogenic mutations

Delivering safer immunotherapies for cancer

Cancer immunotherapy is now a powerful clinical reality, with a steady progression of new drug approvals and a massive pipeline of additional treatments in clinical and preclinical development. However, modulation of the immune system can be a double-edged sword: Drugs that activate immune effectors are prone to serious non-specific systemic inflammation and autoimmune side effects. Drug delivery technologies have an important role to play in harnessing the power of immune therapeutics while avoiding on-target/off-tumor toxicities. Here we review mechanisms of toxicity for clinically-relevant immunotherapeutics, and discuss approaches based in drug delivery technology to enhance the safety and potency of these treatments. These include strategies to merge drug delivery with adoptive cellular therapies, targeting immunotherapies to tumors or select immune cells, and localizing therapeutics intratumorally. Rational design employing lessons learned from the drug delivery and nanomedicine fields has the potential to facilitate immunotherapy reaching its full potential

Die Rolle von Antikörperabhängiger zellulärer Zytotoxizität für die Behandlung des metastatischen kolorektalen Karzinoms mit IgG1 monoklonalen Antikörpern

In the context of cancer therapy, natural killer cells (NK cells) may mediate direct tumour lysis upon binding of their Fcg receptors (FcgR) to IgG1 monoclonal antibodies (mAb) targeted at tumour-expressed surface antigens in a process termed antibody-dependent cellular cytotoxicity (ADCC). Polymorphisms in the FcgRIIIa predominantly expressed by NK cells influence the binding affinity of IgG1 mAbs and in turn their ADCC potential, with the FcgRIIIa-158 V/F single-nucleotide polymorphism being the most extensively studied. In order to elucidate the role of ADCC and FcgRIIIa-158 phenotype in the context of IgG1 mAb therapy, we successfully established a novel setup comprising (1) a flow cytometric FcgRIIIa panel to swiftly determine an individual’s FcgRIIIa-158 phenotype and (2) an experimental model to quantify ADCC activity and immune cell activation in vitro. The FcgRIIIa panel employs an anti-FcgRIIIa clone that only recognises the FcgRIIIa- 158 V allele (MEM-154) in combination with a clone that detects both FcgRIIIa-158 alleles (LNK16). Using the results from melting curve analysis as ground truth, a supervised machine-learning based prediction model computed on the flow cytometry measurements of a training set (n = 39) could be validated with a prediction accuracy > 90% on a test set (n = 52). Importantly, the clinically most relevant distinction between the low- affinity FF phenotype and the high-affinity VF/VV phenotypes was flawless, with the few misclassifications exclusively occurring between the VF and VV phenotypes. Advantages of the FcgRIIIa panel over alternative techniques include its wide applicability, quick turnaround time and low cost. The experimental model based on a 24h co-culture of peripheral blood mononuclear cells (PBMCs) with a range of colorectal cancer cell lines treated with two IgG1 mAbs, namely anti-EGFR cetuximab and/or anti-PDL1 avelumab, could be shown to reliably detect both in vitro ADCC activity and NK cell activation. Experimental readouts were generated using a lactate dehydrogenase release assay together with a flow cytometry panel. In line with the literature, the strength of ADCC activity correlated with the expression (EGFR, PDL1, MICA/B, CD137L) or absence (HLA-A/B/C) of surface markers on tumour cells. These methods were applied to PBMCs derived from 55 patients with RAS and BRAF wild-type metastatic colorectal cancer receiving chemotherapy with cetuximab and avelumab combination therapy within the context of the single-arm multi-centre phase-II FIRE6 clin- ical trial. While FcgRIIIa-158 phenotype and NK cell activation strongly correlated with in vitro ADCC activity, no benefit could be observed for either of these parameters with respect to clinical outcome measures such as response to treatment or progression-free survival.Natürliche Killerzellen (NK Zellen) können direkte Zelllyse mittels des Mechanismus der antikörperabhängigen zellvermittelten Zytotoxizität (engl. ADCC) betreiben, indem sie mit ihren Fcg-Rezeptoren (FcgR) an IgG1 monoklonale Antikörper (mAk) binden, die gegen exprimierte Antigene auf der Oberfläche der Tumorzellen gerichtet sind. Polymorphismen des FcgRIIIa, der sich hauptsächlich auf NK Zellen findet, beeinflussen die Bindungsaffinität für IgG1 mAk und damit das ADCC Potenzial. Unter diesen stellt der FcgRIIIa-158 V/F Einzelnukleotid-Polymorphismus den wissenschaftlich am besten untersuchten dar. Um die Rolle von ADCC und FcgRIIIa-158 Phänotyp für IgG1 mAk-basierte Therapien zu klären, wurde ein experimentelles Setup entwickelt, das aus (1) einem FcgRIIIa Durchflusszytometrie-Assay und (2) einem Modell zur in vitro Quantifizierung der ADCC-Aktivität und Aktivierung von Immunzellpopulationen besteht. Der FcgRIIIa Assay basiert vorrangig auf einem anti-FcgRIIIa Klon (MEM-154), der ausschließlich das FcgRIIIa-158 V Allel erkennt, unter gleichzeitiger Verwendung eines zweiten Klons (LNK16), der beide FcgRIIIa-158 Allele bindet. Aus diesen Messdaten wurde mit Hilfe eines Trainings-Set (n = 39) ein Vorhersagemodell berechnet und anschließend mit einer Genauigkeit von > 90% für ein Test-Set (n = 52) validiert. Dabei gelang die klinisch relevanteste Unterscheidung des FF Phänotyps mit niedriger Bindungsaffinität von den VF und VV Phänotypen mit hoher Bindungsaffinität zweifelsfrei in allen Fällen. Fehlklassifikationen traten ausnahmslos zwischen den Phänotypen VF und VV auf. Die Vorteile des entwickelten FcgRIIIa Assay gegenüber alternativen Methoden bestehen aus einer breiten Anwendbarkeit, kurzer Umlaufzeit und geringen Kosten. Eine zuverlässige Detektion von in vitro ADCC-Aktivität und Aktivierung von NK Zellen konnte durch ein Kokultur-Modell von mononukleären Zellen des peripheren Blutes (engl. PBMCs) mit kolorektalen Krebszelllinien unter Zugabe eines anti-EGFR (Cetuximab) und anti-PDL1 (Avelumab) IgG1 mAk erreicht werden. Experimentelle Werte wurden mittels der Messung von Laktatdehydrogenase-Freisetzung und einem Durchflusszytometrie-Assay quantifiziert. Im Einklang mit der Literatur wurde eine Korrelation zwischen der Stärke der ADCC-Aktivität und der Expression (EGFR, PDL1, MICA/B, CD137L) bzw. des Fehlens (HLA-A/B/C) von Oberflächenmarkern auf den Krebszellen gefunden. Im Rahmen der multizentrischen Phase II FIRE6 Studie wurden die entwickelten Methoden an PBMCs von 55 Patienten mit RAS und BRAF Wildtyp metastatischem kolorektalen Karzinom unter Chemotherapie und Kombinationstherapie von Cetuximab und Avelumab getestet. Obwohl der FcgRIIIa-158 Phänotyp und die Aktivierung von NK Zellen mit der gemessenen in vitro ADCC-Aktivität korrelierten, wurde keine prognostische Relevanz dieser Parameter in Hinblick auf die klinischen Ergebnisgrößen des Therapieansprechens und progressionsfreien Überlebens festgestell