Genetic analysis and natural history of Charcot-Marie-Tooth disease CMTX1 due to GJB1 variants.

Charcot-Marie-Tooth disease (CMT) due to GJB1 variants (CMTX1) is the second most common form of CMT. It is an X-linked disorder characterized by progressive sensory and motor neuropathy with males affected more severely than females. Many reported GJB1 variants remain classified as variants of uncertain significance (VUS). In this large, international, multicentre study we prospectively collected demographic, clinical and genetic data on patients with CMT associated with GJB1 variants. Pathogenicity for each variant was defined using adapted American College of Medical Genetics criteria. Baseline and longitudinal analyses were conducted to study genotype-phenotype correlations, to calculate longitudinal change using the CMT Examination Score (CMTES), to compare males versus females, and pathogenic/likely pathogenic (P/LP) variants versus VUS. We present 387 patients from 295 families harbouring 154 variants in GJB1. Of these, 319 patients (82.4%) were deemed to have P/LP variants, 65 had VUS (16.8%) and three benign variants (0.8%; excluded from analysis); an increased proportion of patients with P/LP variants compared with using ClinVar's classification (74.6%). Male patients (166/319, 52.0%, P/LP only) were more severely affected at baseline. Baseline measures in patients with P/LP variants and VUS showed no significant differences, and regression analysis suggested the disease groups were near identical at baseline. Genotype-phenotype analysis suggested c.-17G>A produces the most severe phenotype of the five most common variants, and missense variants in the intracellular domain are less severe than other domains. Progression of disease was seen with increasing CMTES over time up to 8 years follow-up. Standard response mean (SRM), a measure of outcome responsiveness, peaked at 3 years with moderate responsiveness [change in CMTES (ΔCMTES) = 1.3 ± 2.6, P = 0.00016, SRM = 0.50]. Males and females progressed similarly up to 8 years, but baseline regression analysis suggested that over a longer period, females progress more slowly. Progression was most pronounced for mild phenotypes (CMTES = 0-7; 3-year ΔCMTES = 2.3 ± 2.5, P = 0.001, SRM = 0.90). Enhanced variant interpretation has yielded an increased proportion of GJB1 variants classified as P/LP and will aid future variant interpretation in this gene. Baseline and longitudinal analysis of this large cohort of CMTX1 patients describes the natural history of the disease including the rate of progression; CMTES showed moderate responsiveness for the whole group at 3 years and higher responsiveness for the mild group at 3, 4 and 5 years. These results have implications for patient selection for upcoming clinical trials.

Structure-activity relationships of diverse ACGs against multidrug resistant human lung cancer cell line A549/Taxol.

Fifteen annonaceous acetogenins (ACGs) with different stereochemical structures and configuration, representing three main classes of bis-adjacent-tetrahydrofuran (THF), bis-nonadjacent-THF, and mono-THF ACGs, were selected to tested for their inhibition activity on A549/Taxol cell line, which is multidrug resistant (MDR). The present study showed that some tested compounds showed significant activity toward A549/Taxol cells, and were more potent than the positive control Verapamil. For example, squamostatin-D (14) (IC50 value=16.19µM) was 7.8 times more active than Verapamil (IC50 value=127.09µM). Those ACGs with more carbons between the THF ring and the γ-unsaturated lactone were more potent. Moreover, ACGs with stereochemical arrangement of erythro were more active than those of threo, the compounds with THF ring configuration of cis seemed to be superior to those of trans. However, if all other structural features were identical, the ACGs with more hydroxyls on the aliphatic chain were not more potent towards A549/Taxol, which was not in accordance with previous studies. Furthermore, bis-nonadjacent-THF ACGs whose molecular weight is 622, with three hydroxyl groups located at carbon 16, 19, 24 and stereochemical arrangement of erythro possibly produced notable cytotoxicity. Based on the above conclusions, we proposed a compound model that may be a promising anti-MDR cancer candidate drug in the future clinical trial.

Modelling polycystic liver disease progression using age-adjusted liver volumes and targeted mutational analysis.

Background & Aims: Polycystic liver disease (PLD) manifests as numerous fluid-filled cysts scattered throughout the liver parenchyma. PLD most commonly develops in females, either as an extra-renal manifestation of autosomal-dominant polycystic kidney disease (ADPKD) or as isolated autosomal-dominant polycystic liver disease (ADPLD). Despite known genetic causes, clinical variability challenges patient counselling and timely risk prediction is hampered by a lack of genotype-phenotype correlations and prognostic imaging classifications. Methods: We performed targeted next-generation sequencing and multiplex ligation-dependent probe amplification to identify the underlying genetic defect in a cohort of 80 deeply characterized patients with PLD. Identified genotypes were correlated with total liver and kidney volume (assessed by CT or MRI), organ function, co-morbidities, and clinical endpoints. Results: Monoallelic diagnostic variants were identified in 60 (75%) patients, 38 (48%) of which pertained to ADPKD-gene variants (PKD1, PKD2, GANAB) and 22 (27%) to ADPLD-gene variants (PRKCSH, SEC63). Disease severity defined by age at waitlisting for liver transplantation and first PLD-related hospitalization was significantly more pronounced in mutation carriers compared to patients without genetic diagnoses. While current imaging classifications proved unable to differentiate between severe and moderate courses, grouping by estimated age-adjusted total liver volume progression yielded significant risk discrimination. Conclusion: This study underlines the predictive value of providing a molecular diagnosis for patients with PLD. In addition, we propose a novel risk-classification model based on age- and height-adjusted total liver volume that could improve individual prognostication and personalized clinical management. Lay summary: Polycystic liver disease (PLD) is a highly variable condition that can be asymptomatic or severe. However, it is currently difficult to predict clinical outcomes such as hospitalization, symptom burden, and need for transplantation in individual patients. In the current study, we aimed to investigate the clinical value of genetic confirmation and an age-adjusted total liver volume classification for individual disease prediction. While genetic confirmation generally pointed to more severe disease, estimated age-adjusted increases in liver volume could be useful for predicting clinical outcomes.

Acetogeninas, alternativa en el tratamiento de cáncer en caninos / Acetogenins, alternative cancer treatment in dogs / Acetogeninas, alternativa no tratamento de câncer em caninos

Resumen El cáncer es una enfermedad que implica la alteración de procesos celulares, como metabolismo celular, activación o silenciamiento de genes y crecimiento descontrolado. Es una las principales causas de muerte en humanos, así como en animales de compañía, por lo cual cada vez es más importante la búsqueda y desarrollo de medicamentos. Algunos de los medicamentos que se producen para tratar el cáncer, provienen de plantas, como el taxol y la vincristina. La necesidad de ampliar el uso de productos naturales para tratar esta enfermedad, no solo en humanos sino en animales de compañía como caninos, abre las puertas a la búsqueda de actividad biológica de plantas que son usadas popularmente por presentar algún efecto con la enfermedad. Este es el caso de la guanábana (Annona muricata), de la cual se han extraído metabolitos secundarios (acetogeninas) que presentan in vitro mayor toxicidad en líneas celulares cancerígenas comparadas con líneas celulares normales. El fin de esta revisión fue realizar una aproximación de los usos de estos metabolitos en cáncer, para ello fue hecha una búsqueda en PubMed con diferentes palabras claves y se concluyó, que las acetogeninas comprenden una fuente potencial para el desarrollo de medicamentos contra el cáncer.

Abstract Cancer is a disease that alter cellular processes, like cell metabolism, activation and deactivation of genes and uncontrolled growth. It is one of the main causes of death in humans, as well as in companion animals, which is why it is increasingly important to seek and develop medicines for the treatment. Some of the drugs that are produced to treat cancer come from plants, such as taxol and vincristine. The necessity to expand the use of natural products to treat this disease, not only in humans but also in companion animals such as dogs, opens the doors to seek biological activity of plants that are commonly used because they present some effect over the disease. This is the case of soursop (Annona muricata), from which secondary metabolites (acetogenins) have been extracted and have presented in vitro greater toxicity in cancer cell lines compared to normal cell lines. Therefore, this review was carried out in order to make an approximation of the uses of these metabolites in cancer, for which a search was carried out in PubMed with different key words and it was concluded that acetogenins comprise a potential source for the development of medicines against cancer.

Resumo O câncer é uma doença que envolve a alteração de processos celulares, como o metabolismo celular, ativação ou silenciamento de genes e o crescimento descontrolado. É uma das principais causas de morte em humanos, assim como em animais de companhia, para os quais a busca e o desenvolvimento de medicamentos são cada vez mais importantes. Algumas das drogas que são produzidas para tratar o câncer são derivadas de plantas, como o taxol e a vincristina. A necessidade de ampliar o uso de produtos naturais para tratar esta doença, não somente em humanos, mas em animais de companhia, como cães, abre as portas para a busca da atividade biológica de plantas que são popularmente usadas para causar algum efeito na doença. Este é o caso da graviola (Annona muricata), da qual se extraíram os metabólitos secundários (acetogeninas), que apresentam maior toxicidade in vitro nas linhagens de células cancerígenas do que nas linhagens normais. O objetivo desta revisão foi fazer uma aproximação dos usos desses metabólitos no câncer, para os quais foi feita uma pesquisa em PubMed com palavras-chave diferentes e concluiu-se que as acetogeninas constituem uma fonte potencial para o desenvolvimento de drogas anti-câncer.