Precision oncology in Latin America: current situation, challenges and perspectives.

BACKGROUND: Anti-cancer cytotoxic treatments like platinum-derived compounds often show low therapeutic efficacy, high-risk side effects and resistance. Hence, targeted treatments designed to attack only tumour cells avoiding these harmful side effects are highly needed in clinical practice. Due to this, precision oncology has arisen as an approach to specifically target alterations present only in cancer cells, minimising side effects for patients. It involves the use of molecular biomarkers present in each kind of tumour for diagnosis, prognosis and treatment. Since these biomarkers are specific for each cancer type, physicians use them to stratify, diagnose or take the best therapeutic options for each patient depending on the features of the specific tumour. AIM: This review aims to describe the current situation, limitations, advantages and perspectives about precision oncology in Latin America. MAIN BODY: For many years, many biomarkers have been used in a clinical setting in developed countries. However, in Latin American countries, their broad application has not been affordable partially due to financial and technical limitations associated with precarious health systems and poor access of low-income populations to quality health care. Furthermore, the genetic mixture in Latin American populations could generate differences in treatment responses from one population to another (pharmacoethnicity) and this should be evaluated before establishing precision therapy in particular populations. Some research groups in the region have done a lot of work in this field and these data should be taken as a starting point to establish networks oriented to finding clinically useful cancer biomarkers in Latin American populations. CONCLUSION: Latin America must create policies allowing excluded populations to gain access to health systems and next generation anti-cancer drugs, i.e. high-cost targeted therapies to improve survival. Also, cancer clinical research must be oriented to establish cancer biomarkers adapted to specific populations with different ethnicity, allowing the improvement of patient outcomes.

Hypoxic resistance of KRAS mutant tumor cells to 3-Bromopyruvate is counteracted by Prima-1 and reversed by N-acetylcysteine.

BACKGROUND: The metabolic inhibitor 3-bromopyruvate (3-BrPA) is a promising anti-cancer alkylating agent, shown to inhibit growth of some colorectal carcinoma with KRAS mutation. Recently, we demonstrated increased resistance to 3-BrPA in wt p53 tumor cells compared to those with p53 silencing or mutation. Since hypoxic microenvironments select for tumor cells with diminished therapeutic response, we investigated whether hypoxia unequally increases resistance to 3-BrPA in wt p53 MelJuso melanoma harbouring (Q61L)-mutant NRAS and wt BRAF, C8161 melanoma with (G12D)-mutant KRAS (G464E)-mutant BRAF, and A549 lung carcinoma with a KRAS (G12S)-mutation. Since hypoxia increases the toxicity of the p53 activator, Prima-1 against breast cancer cells irrespective of their p53 status, we also investigated whether Prima-1 reversed hypoxic resistance to 3-BrPA. RESULTS: In contrast to the high susceptibility of hypoxic mutant NRAS MelJuso cells to 3-BrPA or Prima-1, KRAS mutant C8161 and A549 cells revealed hypoxic resistance to 3-BrPA counteracted by Prima-1. In A549 cells, Prima-1 increased p21CDKN1mRNA, and reciprocally inhibited mRNA expression of the SLC2A1-GLUT1 glucose transporter-1 and ALDH1A1, gene linked to detoxification and stem cell properties. 3-BrPA lowered CAIX and VEGF mRNA expression. Death from joint Prima-1 and 3-BrPA treatment in KRAS mutant A549 and C8161 cells seemed mediated by potentiating oxidative stress, since it was antagonized by the anti-oxidant and glutathione precursor N-acetylcysteine. CONCLUSIONS: This report is the first to show that Prima-1 kills hypoxic wt p53 KRAS-mutant cells resistant to 3-BrPA, partly by decreasing GLUT-1 expression and exacerbating pro-oxidant stress.

Optimized Multiplex Detection of 7 KRAS Mutations by Taqman Allele-Specific qPCR.

UNLABELLED: Establishing the KRAS mutational status of tumor samples is essential to manage patients with colorectal or lung cancer, since these mutations preclude treatment with monoclonal anti-epidermal growth factor receptor (EGFR) antibodies. We report an inexpensive, rapid multiplex allele-specific qPCR method detecting the 7 most clinically relevant KRAS somatic mutations with concomitant amplification of non-mutated KRAS in tumor cells and tissues from CRC patients. Positive samples evidenced in the multiplex assay were further subjected to individual allele-specific analysis, to define the specific mutation. Reference human cancer DNA harbouring either G12A, G12C, G12D, G12R, G12S, G12V and G13D confirmed assay specificity with ≤1% sensitivity of mutant alleles. KRAS multiplex mutation analysis usefulness was also demonstrated with formalin-fixed paraffin embedded (FFPE) from CRC biopsies. CONCLUSION: Co-amplification of non-mutated DNA avoided false negatives from degraded samples. Moreover, this cost effective assay is compatible with mutation detection by DNA sequencing in FFPE tissues, but with a greater sensitivity when mutant DNA concentrations are limiting.

Identificación de una secuencia de ADN genómico de Leishmania especifica del subgénero Leishmania / Identification of a genomic DNA sequence of Leishmania, specific of Leishmania subgenus

Leishmania es el agente causante de la compleja enfermedad conocida como leishmaniasis. Las distintas especies de este parásito protozoario se encuentran agrupadas en dos subgéneros, Viannia y Leishmania, de acuerdo a su desarrollo en el mosquito vector. Un ensayo de PCR, β500-PCR, específico del subgénero Viannia, ha sido desarrollado utilizando la secuencia de ADN genómico denominada β500. En este trabajo se presenta el aislamiento e identificación de una secuencia genómica de 280 pb, L280, a partir del ADN genómico de Leishmania (Leishmania) mexicana luego de aplicar el ensayo β500-PCR en condiciones de baja rigurosidad. La secuenciación parcial de L280 permitió diseñar un ensayo de PCR (L280-PCR) que generó un producto de amplificación de 260 pb, en distintas condiciones de rigurosidad, cuando se utilizó el ADN genómico de distintas especies pertenecientes al subgénero Leishmania. El ensayo L280-PCR resultó negativo para el ADN genómico de distintas especies del subgénero Viannia al igual que para el ADN de otros organismos kinetoplastidos o humano. Los resultados sugieren que el ensayo L280-PCR es específico del subgénero Leishmania.

Leishmania is the causal agent of the leishmaniasis disease. The different species of this protozoa parasite are grouped in two subgenera, Viannia and Leishmania, according to their development in the sandfly vector. A specific PCR assay, β500-PCR, has been developed for the Viannia subgenus using the genomic β500 DNA sequence. In the present work we present the isolation and identification of a genomic sequence of 280 bp, L280, obtained from genomic DNA of Leishmania (Leishmania) mexicana after application of the β500-PCR assay at low stringency. After partial sequencing of L280 a PCR assay was generated, L280-PCR, this yielded a product of 260 bp at different conditions of stringency, when genomic DNA of different species of Leishmania subgenus was used. The L280-PCR assay was negative to genomic DNA of species belonging to the Viannia subgenus and also to other kinetoplastid organisms and human. The results suggest specificity of the L280-PCR assay for the Leishmania subgenus.