Protein Aggregation and Protein Instability Govern Familial Amyotrophic Lateral Sclerosis Patient Survival

The nature of the “toxic gain of function” that results from amyotrophic lateral sclerosis (ALS)-, Parkinson-, and Alzheimer-related mutations is a matter of debate. As a result no adequate model of any neurodegenerative disease etiology exists. We demonstrate that two synergistic properties, namely, increased protein aggregation propensity (increased likelihood that an unfolded protein will aggregate) and decreased protein stability (increased likelihood that a protein will unfold), are central to ALS etiology. Taken together these properties account for 69% of the variability in mutant Cu/Zn-superoxide-dismutase-linked familial ALS patient survival times. Aggregation is a concentration-dependent process, and spinal cord motor neurons have higher concentrations of Cu/Zn-superoxide dismutase than the surrounding cells. Protein aggregation therefore is expected to contribute to the selective vulnerability of motor neurons in familial ALS

Circulating regulatory T cells from breast cancer patients in response to neoadjuvant chemotherapy

Background: Immune escape of tumor cells is a new hallmark of cancer in general, and breast cancer, in particular. Previous studies have demonstrated that the immunological profile in peripheral blood may be a prognostic and/or predictive biomarker in breast cancer. Thus, higher number of regulatory T cells (Tregs) in blood from patients with breast cancer has been reported in relation to normal donors. In the present study, we planned to evaluate the changes in different cell populations in peripheral blood: neutrophils, monocytes and lymphocytes, as well as lymphocyte subpopulations [natural killer (NK), B lymphocytes, T lymphocytes, both CD4(+) and CD8(+), and Tregs] from patients with local breast cancer (both Her2(+) and Her2(-)), before, during and after neoadjuvant chemotherapy.Methods: We have employed flow cytometry for the cell analysis of fresh samples obtained before and whilst the neoadjuvant treatment was accomplished. We have studied 50 successive patients from the Breast Cancer Unit of the Virgen Macarena University Hospital during 2 years.Results: Neoadjuvant chemotherapy induced a significant reduction in B cells, especially in Her2(-) patients, and a reduction in NK cells. CD4(+) T cells decreased, whereas CD8(+) cells only decreased in Her2(-) patients. Tregs were also diminished, especially in Her2(+) patients, in response to treatment. Thus, higher CD8/Treg ratio was observed in Her2(+) patients. A higher percentage of Her2(+) patients (66.6%) achieved complete response than Her2(-) patients (27.5%). Monocytes and neutrophils were not changed in peripheral blood.Conclusions: Even though the decrease in B cells and NK cells in response to chemotherapy may be deleterious in the neoadjuvant treatment of breast cancer, the decrease in Tregs and CD4 T cells, but not CD8 T cells, increasing the CD8/Treg ratio, especially in Her2(+) patients, may reveal a new tool to monitor the immune response in breast cancer treated with chemotherapy in the neoadjuvant setting

Circulating myeloid-derived suppressor cells and regulatory T cells as immunological biomarkers in refractory/relapsed diffuse large B-cell lymphoma: translational results from the R2-GDP-GOTEL trial

Background: The search for immunological markers with ability of predicting clinical outcome is a priority in lymphomas, and in cancer in general. It is well known that some immunomodulatory cells, such as myeloid derived suppressor cells (MDSCs) or regulatory T cells (Tregs), are recruited by tumors, jeopardizing antitumor immunosurveillance. In this work, we have studied blood levels of these immunosuppressive cells in patients with relapsed/refractory diffuse large B-cell lymphoma (R/R DLBCL), prior to and along the course of the experimental rituximab, gemcitabine, dexamethasone, and cisplatin (R2-GDP) schedule, as a translational substudy of the R2-GDP-GOTEL trial (EudraCT Number: 2014-001620-29), which included lenalidomide as an immunomodulator. Methods: Blood samples were taken before treatment, at cycle 3 and end of induction. Samples were analyzed by flow cytometry. Non-parametric tests were used. Mann-Whitney U test was used to compare basal cells distributions, and Wilcoxon test was considered to compare cells distribution at different times. Spearman test was performed to measure the degree of association between cell populations. Results: In this study, MDSC and Treg circulating concentration was found increased in all patients compared with a healthy control group and decreased after treatment only in patients with longest overall survival (>24 months), reaching the levels of the healthy group. Likewise, the number of inhibited T lymphocytes expressing Programmed Death-1 (PD-1) were increased in peripheral blood from patients and decreased on the treatment, whereas activated T lymphocytes increased after therapy in those with better overall survival. Conclusions: In conclusion, blood concentration of MDSCs and Treg cells may be good prognostic markers for overall survival after 2 years in R/R DLBCL. These results point to a possible role of these elements in the immunosuppression of these patients, as assessed by the circulating activated and inhibited T lymphocytes, and therefore, they may be considered as therapeutic targets in DLBCL

DPY19L2, its mutation in about half globozoospermia

International audienceIntroduction: Testicular sperm extraction (TESE) combined with intracytoplasmic sperm injection is a promising assessment in reproductive practice particularly for patients with non obstructive azoospermia (NOA). There was evidence that impaired spermatogenesis could be related to an imbalance in the intratesticular oestradiol/testosterone ratio. We carried out a prospective observational study in order to evaluate the putative variation of the expression of genes implicated in the estrogen synthesis (aromatase) and mediation of estrogen action (estrogen receptors and GRP30 for the respective initiation of genomic and non-genomic pathways) in human testicular biopsies and to understand the mechanisms involved in different testicular disorders in relation to NOA such as hypospermatogenesis (Hsg), germ cell arrest (GCA) and Sertoli Cell Only (SCO) syndrome. Material and Methods: Histological evaluation, sperm recovery and ARN extraction followed by the measurement of relative mRNA level of cyp19, Esr1, Esr2 and gpr30 using real time polymerase chain reaction were realized in testicular bilateral fragments (n = 98) providing from 49 azoospermic patients. Taking into account the existence of potential discordant patterns in bilateral biopsies, the high prevalence of mixed patterns in a same testes and the fact that histological evaluation was always performed in a testicular biopsy different from this studied, we have reported the expression of specific genes considered as cells markers (Prm1 for round spermatids, H1t for pachytene spermatocytes and vimentin for mature Sertoli cells) for the selection of pure and homogeneous NOA forms. Then the expression of genes encoding for aromatase, estrogen receptors and GPR 30 has been evaluated in obstructive azoospermia group (0A) used as control and NOA groups (Hsg,GCA and SCO). Results are expressed as means + S.E.M. Statistical analysis was performed using ANOVA (Graphpad Instat 3, GraphPad Software, San Diego, CA, USA) and means are compared using Tukey-Kramer multiple comparisons test. Statistical significance was accepted at p < 0.05. Results: We have at first described specific patterns of pure forms of Hsg, GCA, SCO and OA with the helpful of cell markers. A pure form of SCO could be defined as a relative expression of vimentin transcript higher than 2 associated with an absence of Prm 1 or H1t transcripts. The level of Prm1 transcripts and the ratio Prm1 mRNA/H1t mRNA are significantly correlated with the number of spermatozoa recovered by TESE. A reduced expression of GRP30 is observed in all groups but seems more elevated in GCA group. Levels of the two isoforms ERalpha and ERbeta transcripts are significantly increased in OA and Hsg groups. But only the ERalpha level is strongly correlated with that of Prm1 and sperm recovery. Aromatase expression doest not differ significantly in the four groups studied. However we have found an intensive expression of aromatase and ERalpha in the SCO group associated with Leydig cell hyperplasia. Conclusions: Studying the putative variation of transcripts implicated in the estrogen synthesis and mediation of estrogen action in testicular biopsies could represent a helpful for the understanding of mechanisms involved in the pathogenesis of NOA forms and bring new insights about the role of estrogens during spermatogenesis. GRP30 expression seems to be restricted to testicular cells implicated in the first steps of spermatogenesis. The relative important expression of the two isoforms ERalpha and ERbeta in post-meiotic cells suggests their role during spermiogenesis. But an enhanced expression of ERalpha in Leydig cell hyperplasia and a tight correlation between ERalpha and Prm1 expression could argue the case for a differential role of the two ER isoforms during spermatogenesis