High-throughput analysis of the RNA-induced silencing complex in myotonic dystrophy type 1 patients identifies the dysregulation of miR-29c and its target ASB2

Myotonic dystrophy type 1 (DM1) is a multi-systemic disorder caused by abnormally expanded stretches of CTG DNA triplets in the DMPK gene, leading to mutated-transcript RNA-toxicity. MicroRNAs (miRNAs) are short non-coding RNAs that, after maturation, are loaded onto the RISC effector complex that destabilizes target mRNAs and represses their translation. In DM1 muscle biopsies not only the expression, but also the intracellular localization of specific miRNAs is disrupted, leading to the dysregulation of the relevant mRNA targets. To investigate the functional alterations of the miRNA/target interactions in DM1, we analyzed by RNA-sequencing the RISC-associated RNAs in skeletal muscle biopsies derived from DM1 patients and matched controls. The mRNAs found deregulated in DM1 biopsies were involved in pathways and functions relevant for the disease, such as energetic metabolism, calcium signaling, muscle contraction and p53-dependent apoptosis. Bioinformatic analysis of the miRNA/mRNA interactions based on the RISC enrichment profiles, identified 24 miRNA/mRNA correlations. Following validation in 21 independent samples, we focused on the couple miR-29c/ASB2 because of the role of miR-29c in fibrosis (a feature of late-stage DM1 patients) and of ASB2 in the regulation of muscle mass. Luciferase reporter assay confirmed the direct interaction between miR-29c and ASB2. Moreover, decreased miR-29c and increased ASB2 levels were verified also in immortalized myogenic cells and primary fibroblasts, derived from biopsies of DM1 patients and controls. CRISPR/Cas9-mediated deletion of CTG expansions rescued normal miR-29c and ASB2 levels, indicating a direct link between the mutant repeats and the miRNA/target expression. In conclusion, functionally relevant miRNA/mRNA interactions were identified in skeletal muscles of DM1 patients, highlighting the dysfunction of miR-29c and ASB2

Influencia de los Microplásticos (MPs) y Productos Farmacéuticos y de Cuidado Personal (PPCPs) Derivados de la Actividad Humana Costera y sus Efectos en el Ecosistema Antártico.

Tesis presentada para optar al grado de Doctor en Ciencias Ambientales con mención en Sistemas Acuáticos ContinentalesEl incremento de la actividad humana en la Antártica ha generado numerosos impactos en el ecosistema. Recientemente ha sido descrita la presencia de microplásticos (MPs) y Productos farmacéuticos y de cuidado personal (PPCPs) en ambientes costeros, principalmente en la Península Antártica. El objetivo de esta tesis fue evaluar la contribución de la actividad humana en las concentraciones de MPs y PPCPs en el ecosistema antártico. Para esto, se determinó la abundancia y composición de microplásticos en suelos y en sedimentos intermareales de la Bahía Fildes, Antártica. En los suelos se obtuvo un promedio de 13,6 partículas/50 ml de muestra, con una longitud máxima de partículas de <500 µm. Estas partículas fueron identificadas como fragmentos, caracterizados por su composición basada en resina fenoxi de color naranjo, idénticos a los revestimientos de las instalaciones de la base Frei. En contraste, en los sedimentos intermareales se registraron exclusivamente fibras en una menor abundancia (1,5 partículas/50 ml de muestra), pero mayor longitud (<2000 µm). Estas fibras se caracterizaron po una composición de tereftalato de polietileno (PET) de distintos colores, con mayores abundancias en los sitios cercanos a los efluentes de aguas residuales, donde además se encontraron fibras de algodón, lo que permite presumir que su principal fuente corresponde a aguas grises. Respecto a las variables ambientales evaluadas, si bien la materia orgánica se correlacionó positivamente con los fragmentos de plástico del suelo, esta variable parece explicarse más en la composición de los microplásticos. En tanto, las fibras de los sedimentos intermareales se correlacionaron positivamente al tamaño de partícula de los sedimentos. En general, las variables ambientales seleccionadas en este estudio parecen no explicar las concentraciones de microplásticos. Finalmente, se observó que, a pesar de las restricciones impuestas en zonas protegidas, la contaminación por microplásticos se ha expandido a éstas áreas. Un objetivo adicional fue identificar las fuentes, concentraciones y distribución de PPCPs en el sistema costero antártico y sus efectos en la biota. De modo que se realizó una revisión de los artículos científicos disponibles en la Antártica, junto a la bibliografía del Consejo de Administradores de Programas Antárticos Nacionales (CONMAP) y la Secretaría del Tratado Antártico (ATS). Esto se complementó con los datos espaciales de Quantarctica. Los resultados muestran que existe una amplia variedad de PPCPs reportados en aguas costeras los que alcanzan concentraciones de hasta 60000 ng/l. Las mayores concentraciones se detectaron en base Esperanza debido a la ausencia de tratamiento, siendo significativamente elevados para los analgésicos acetaminofén (48744 ng/l), diclofenaco (15087 ng/l) e ibuprofeno (10053 ng/l). No obstante, la implementación de un sistema de tratamiento terciario en base Scott igualmente arrojó altos valores para los filtros UV 4- metil-bencilidenoalcanfor (4-MBC) (<11700 ng/l) y 2,4- dihidroxibenzofenona (BP-1) (<6830 ng/l), así como el surfactante 4-t-octilfenol (OP) (<7050 ng/l). En general, se observó que el número de ocupantes de cada estación no fue tan determinante como la implementación de algún sistema de tratamiento de aguas residuales. Sin embargo, otras variables como la frecuencia de muestreo también deben ser consideradas. De este modo, se identificó a los efluentes como la principal fuente de PPCPs en la Antártica, siendo mayores las concentraciones en zonas aledañas a las bases en comparación de mar adentro. Además, la biota antártica ha sido poco estudiada, con solo una especie evaluada, por lo tanto, los eventuales efectos de los microplásticos son por ahora desconocidos. Esta tesis demuestra que las actividades humanas en la Antártica han resultado en la contaminación por MPs y PPCPs de sus suelos, agua de mar y sedimentos costeros, y que existe un alto grado de desconocimiento de los eventuales efectos en la biota. Dado que la mayoría de las estaciones antárticas se localizan en las zonas costeras, sus efluentes tienen una gran influencia sobre el ambiente marino, siendo identificados como la principal fuente de contaminación por MPs y PPCPs. En contraste, los suelos son dominados por la contaminación de microplásticos que en bahía Fildes responde al uso de suelo mediante la infraestructura. En este sentido, debido a las restricciones que rigen del Protocolo Ambiental, las limitadas actividades efectuadas dentro del territorio antártico sólo permitirían el uso y posterior eliminación de PPCPs principalmente mediante los efluentes, de modo que su presencia en suelos se espera que sea escasa o nula. El principal inconveniente es que algunas de las estaciones de investigación carecen de tratamiento terciario, descrito como uno de los más efectivos en la remoción de PPCPs. Asimismo, se permite la liberación de efluentes sin tratamiento directamente al mar. En consecuencia, mayores esfuerzos por sobre lo dictado por el Protocolo Ambiental, como el mejoramiento voluntario de estos sistemas de tratamiento y el control, tanto en el ingreso como uso de estos contaminantes, son medidas que podrían compensar la deficiente remoción de algunos contaminantes, ante la espera de nuevas regulaciones de los contaminantes emergentes antes mencionados.The increase in human activity in Antarctica has generated numerous impacts on the ecosystem. The presence of microplastics (MPs) and pharmaceutical and personal care products (PPCPs) in coastal environments, mainly in the Antarctic Peninsula, has recently been described. The objective of this thesis was to evaluate the contribution of human activity in the concentrations of PMs and PPCPs in the Antarctic ecosystem. For this, the abundance and composition of microplastics in soils and intertidal sediments of Fildes Bay, Antarctica, were determined. In the soils, an average of 13.6 particles/50 ml of sample was obtained, with a maximum particle length of <500 µm. These particles were identified as fragments, characterized by their composition based on orange-colored phenoxy resin, identical to the coatings of the Frei base facilities. In contrast, in the intertidal sediments fibers were exclusively recorded in a lower abundance (1.5 particles/50 ml of sample), but greater length ( <2000 µm). These fibers were characterized by a composition of polyethylene terephthalate (PET) of different colors, with greater abundances in the sites near the wastewater effluents, where cotton fibers were also found, which allows us to presume that their main source corresponds to grey waters. Regarding the environmental variables evaluated, although organic matter was positively correlated with plastic fragments in the soil, this variable seems to be explained more by the composition of microplastics. Meanwhile, intertidal sediment fibers were positively correlated to sediment particle size. In general, the environmental variables selected in this study do not seem to explain the concentrations of microplastics. Finally, it was observed that, despite the restrictions imposed in protected areas, microplastic contamination has spread to these sites. An additional objective was to identify the sources, concentrations and distribution of PPCPs in the Antarctic coastal system and their effects on biota. A review of the scientific articles available in Antarctica was carried out, together with the bibliography of the Council of Administrators of National Antarctic Programs (CONMAP) and the Secretariat of the Antarctic Treaty (ATS). This was supplemented by spatial data from Quantarctica. The results show that there is a wide variety of PPCPs reported in coastal waters, reaching concentrations of up to 60,000 ng/l. The highest concentrations were detected in the Esperanza base due to the absence of treatment, being significantly high for the analgesics acetaminophen (48744 ng/l), diclofenac (15087 ng/l) and ibuprofen (10053 ng/l). However, the implementation of a Scott-based tertiary treatment system also yielded high values for the UV filters 4-methyl benzylidenecamphor (4-MBC) (<11700 ng/l) and 2,4-dihydroxybenzophenone (BP-1) (<6830 ng/l), as well as the surfactant 4-t-octylphenol (OP) (<7050 ng/l). In general, it was observed that the number of occupants of each station was not as decisive as the implementation of a wastewater treatment system. However, other variables such as sampling frequency must also be considered. In this way, effluents were identified as the main source of PPCPs in Antarctica, with higher concentrations in areas surrounding the bases compared to offshore. In addition, the Antarctic biota has been little studied, with only one species evaluated, therefore, the eventual effects of microplastics are unknown at this time. This thesis demonstrates that human activities in Antarctica have resulted in contamination by PMs and PPCPs of its soils, seawater and coastal sediments, and little is known about the possible effects on the biota. Since most Antarctic stations are located in coastal areas, their effluents have a great influence on the marine environment, being identified as the main source of pollution by PMs and PPCPs. In contrast, the soils are dominated by microplastic contamination that in Fildes Bay responds to land use through infrastructure. In this sense, due to the restrictions that govern the Environmental Protocol, the limited activities carried out within the Antarctic territory would only allow the use and subsequent elimination of PPCPs, mainly through effluents, so that their presence in soils is expected to be scarce or null. The main drawback is that some of the research stations lack tertiary treatment, described as one of the most effective in removing PPCPs. Likewise, the release of effluents without treatment directly into the sea is allowed. Consequently, greater efforts beyond what is dictated by the Environmental Protocol, such as the voluntary improvement of these treatment systems and the control, both in the entry and use of these pollutants, are measures that could compensate for the poor removal of some pollutants, given waiting for new regulations of the aforementioned emerging pollutants.ANID, Beca Apoyo de Tesis Doctoral N° 24110019 ANID-PFCHA/Doctorado Nacional/2017-21170746Millennium Science Initiative Program, Código ICN2019_015Facultad de Ciencias AmbientalesDepartamento de Sistemas AcuáticosConcepció

Deregulated MicroRNAs in Myotonic Dystrophy Type 2

Myotonic Dystrophy Type-2 (DM2) is an autosomal dominant disease caused by the expansion of a CCTG tetraplet repeat. It is a multisystemic disorder, affecting skeletal muscles, the heart, the eye, the central nervous system and the endocrine system. Since microRNA (miRNA) expression is disrupted in Myotonic Dystrophy Type-1 and many other myopathies, miRNAs deregulation was studied in skeletal muscle biopsies of 13 DM2 patients and 13 controls. Eleven miRNAs were deregulated: 9 displayed higher levels compared to controls (miR-34a-5p, miR-34b-3p, miR-34c-5p, miR-146b-5p, miR-208a, miR-221-3p and miR-381), while 4 were decreased (miR-125b-5p, miR-193a-3p, miR-193b-3p and miR-378a-3p). To explore the relevance of DM2 miRNA deregulation, the predicted interactions between miRNA and mRNA were investigated. Global gene expression was analyzed in DM2 and controls and bioinformatic analysis identified more than 1,000 miRNA/mRNA interactions. Pathway and function analysis highlighted the involvement of the miRNA-deregulated mRNAs in multiple aspects of DM2 pathophysiology. In conclusion, the observed miRNA dysregulations may contribute to DM2 pathogenetic mechanisms

Association of Upfront Peptide Receptor Radionuclide Therapy With Progression-Free Survival Among Patients With Enteropancreatic Neuroendocrine Tumors

open57noIMPORTANCE Data about the optimal timing for the initiation of peptide receptor radionuclide therapy (PRRT) for advanced, well-differentiated enteropancreatic neuroendocrine tumors are lacking. OBJECTIVE To evaluate the association of upfront PRRT vs upfront chemotherapy or targeted therapy with progression-free survival (PFS) among patients with advanced enteropancreatic neuroendocrine tumors who experienced disease progression after treatment with somatostatin analogues (SSAs). DESIGN, SETTING, AND PARTICIPANTS This retrospective, multicenter cohort study analyzed the clinical records from 25 Italian oncology centers for patients aged 18 years or older who had unresectable, locally advanced or metastatic, well-differentiated, grades 1 to 3 enteropancreatic neuroendocrine tumors and received either PRRT or chemotherapy or targeted therapy after experiencing disease progression after treatment with SSAs between January 24, 2000, and July 1, 2020. Propensity score matching was done to minimize the selection bias. EXPOSURES Upfront PRRT or upfront chemotherapy or targeted therapy. MAIN OUTCOMES AND MEASURES The main outcome was the difference in PFS among patients who received upfront PRRT vs among those who received upfront chemotherapy or targeted therapy. A secondary outcome was the difference in overall survival between these groups. Hazard ratios (HRs) were fitted in a multivariable Cox proportional hazards regression model to adjust for relevant factors associated with PFS and were corrected for interaction with these factors. RESULTS Of 508 evaluated patients (mean ([SD] age, 55.7 [0.5] years; 278 [54.7%] were male), 329 (64.8%) received upfront PRRT and 179 (35.2%) received upfront chemotherapy or targeted therapy. The matched group included 222 patients (124 [55.9%] male; mean [SD] age, 56.1 [0.8] years), with 111 in each treatment group. Median PFS was longer in the PRRT group than in the chemotherapy or targeted therapy group in the unmatched (2.5 years [95%CI, 2.3-3.0 years] vs 0.7 years [95%CI, 0.5-1.0 years]; HR, 0.35 [95%CI, 0.28-0.44; P &lt; .001]) and matched (2.2 years [95% CI, 1.8-2.8 years] vs 0.6 years [95%CI, 0.4-1.0 years]; HR, 0.37 [95%CI, 0.27-0.51; P &lt; .001]) populations. No significant differences were shown in median overall survival between the PRRT and chemotherapy or targeted therapy groups in the unmatched (12.0 years [95%CI, 10.7-14.1 years] vs 11.6 years [95%CI, 9.1-13.4 years]; HR, 0.81 [95%CI, 0.62-1.06; P = .11]) and matched (12.2 years [95% CI, 9.1-14.2 years] vs 11.5 years [95%CI, 9.2-17.9 years]; HR, 0.83 [95%CI, 0.56-1.24; P = .36]) populations. The use of upfront PRRT was independently associated with improved PFS (HR, 0.37; 95%CI, 0.26-0.51; P &lt; .001) in multivariable analysis. After adjustment of values for interaction, upfront PRRT was associated with longer PFS regardless of tumor functional status (functioning: adjusted HR [aHR], 0.39 [95%CI, 0.27-0.57]; nonfunctioning: aHR, 0.29 [95%CI, 0.16-0.56]), grade of 1 to 2 (grade 1: aHR, 0.21 [95%CI, 0.12-0.34]; grade 2: aHR, 0.52 [95%CI, 0.29-0.73]), and site of tumor origin (pancreatic: aHR, 0.41 [95%CI, 0.24-0.61]; intestinal: aHR, 0.19 [95%CI, 0.11-0.43]) (P &lt; .001 for all). Conversely, the advantage was not retained in grade 3 tumors (aHR, 0.31; 95%CI, 0.12-1.37; P = .13) or in tumors with a Ki-67 proliferation index greater than 10% (aHR, 0.73; 95%CI, 0.29-1.43; P = .31). CONCLUSIONS AND RELEVANCE In this cohort study, treatment with upfront PRRT in patients with enteropancreatic neuroendocrine tumors who had experienced disease progression with SSA treatment was associated with significantly improved survival outcomes compared with upfront chemotherapy or targeted therapy. Further research is needed to investigate the correct strategy, timing, and optimal specific sequence of these therapeutic options.openPusceddu, Sara; Prinzi, Natalie; Tafuto, Salvatore; Ibrahim, Toni; Filice, Angelina; Brizzi, Maria Pia; Panzuto, Francesco; Baldari, Sergio; Grana, Chiara M.; Campana, Davide; Davì, Maria Vittoria; Giuffrida, Dario; Zatelli, Maria Chiara; Partelli, Stefano; Razzore, Paola; Marconcini, Riccardo; Massironi, Sara; Gelsomino, Fabio; Faggiano, Antongiulio; Giannetta, Elisa; Bajetta, Emilio; Grimaldi, Franco; Cives, Mauro; Cirillo, Fernando; Perfetti, Vittorio; Corti, Francesca; Ricci, Claudio; Giacomelli, Luca; Porcu, Luca; Di Maio, Massimo; Seregni, Ettore; Maccauro, Marco; Lastoria, Secondo; Bongiovanni, Alberto; Versari, Annibale; Persano, Irene; Rinzivillo, Maria; Pignata, Salvatore Antonio; Rocca, Paola Anna; Lamberti, Giuseppe; Cingarlini, Sara; Puliafito, Ivana; Ambrosio, Maria Rosaria; Zanata, Isabella; Bracigliano, Alessandra; Severi, Stefano; Spada, Francesca; Andreasi, Valentina; Modica, Roberta; Scalorbi, Federica; Milione, Massimo; Sabella, Giovanna; Coppa, Jorgelina; Casadei, Riccardo; Di Bartolomeo, Maria; Falconi, Massimo; de Braud, FilippoPusceddu, Sara; Prinzi, Natalie; Tafuto, Salvatore; Ibrahim, Toni; Filice, Angelina; Brizzi, Maria Pia; Panzuto, Francesco; Baldari, Sergio; Grana, Chiara M.; Campana, Davide; Davì, Maria Vittoria; Giuffrida, Dario; Zatelli, Maria Chiara; Partelli, Stefano; Razzore, Paola; Marconcini, Riccardo; Massironi, Sara; Gelsomino, Fabio; Faggiano, Antongiulio; Giannetta, Elisa; Bajetta, Emilio; Grimaldi, Franco; Cives, Mauro; Cirillo, Fernando; Perfetti, Vittorio; Corti, Francesca; Ricci, Claudio; Giacomelli, Luca; Porcu, Luca; Di Maio, Massimo; Seregni, Ettore; Maccauro, Marco; Lastoria, Secondo; Bongiovanni, Alberto; Versari, Annibale; Persano, Irene; Rinzivillo, Maria; Pignata, Salvatore Antonio; Rocca, Paola Anna; Lamberti, Giuseppe; Cingarlini, Sara; Puliafito, Ivana; Ambrosio, Maria Rosaria; Zanata, Isabella; Bracigliano, Alessandra; Severi, Stefano; Spada, Francesca; Andreasi, Valentina; Modica, Roberta; Scalorbi, Federica; Milione, Massimo; Sabella, Giovanna; Coppa, Jorgelina; Casadei, Riccardo; Di Bartolomeo, Maria; Falconi, Massimo; de Braud, Filipp

Case Report: Successful use of emapalumab in adult B-cell acute lymphoblastic leukemia experiencing severe neurotoxicity and hemophagocytic lymphohistiocytosis-like features after CAR-T cell therapy

Chimeric antigen receptor (CAR)-T cell therapy is a powerful adoptive immunotherapy associated with significant toxicity, including cytokine release syndrome (CRS) and immune effector cell-associated neurotoxicity syndrome (ICANS). As CAR-T usage expands, hyperinflammatory toxicities resembling hemophagocytic lymphohistiocytosis (HLH) syndrome are increasingly recognized. Immune effector cell-associated HLH-like syndrome (IEC-HS) describes HLH-like symptoms attributable to CAR-T cell therapy, often presenting as CRS resolves. Treatments for IEC-HS are adapted from primary HLH, including corticosteroids, the recombinant human interleukin (IL)-1 receptor antagonist anakinra and the Janus Kinase inhibitor ruxolitinib. Emapalumab, an anti-IFN-γ antibody, is promising but underexplored in adult IEC-HS cases. We report an adult B-cell acute lymphoblastic leukemia (B-ALL) patient treated with brexucabtagene autoleucel (brexu-cel). The patient developed CRS, refractory neurotoxicity, and IEC-HS with worsening multiorgan failure and hyperinflammatory markers. Treatment included tocilizumab, high-dose corticosteroids, anakinra, siltuximab, and ruxolitinib. Despite aggressive management, hyperinflammation and neurotoxicity persisted. Emapalumab was initiated on day +11, resulting in normalization of the biochemical parameters and full neurological recovery by day +21. The patient recovered from IEC-HS and underwent allogeneic stem cell transplantation. This case highlights the role of emapalumab in managing refractory IEC-HS and persistent neurotoxicity in adults, underscoring the need for targeted interventions in severe CAR-T complications

Plomo en suelos de colonias de pingüinos del género Pygoscelis en la Península Ardley, Antártica.

Seminario para optar al título de Biólogo.Los metales pesados son elementos químicos que en determinadas concentraciones benefician a los organismos, pero al encontrarse en concentraciones anómalas en el ambiente debido a causas naturales o antropogénicas, ingresan al agua donde su elevada persistencia en el medio les otorga la capacidad de biomagnificarse y bioacumularse en las cadenas tróficas, resultando tóxicos para las células. El plomo se encuentra naturalmente en la corteza terrestre y su principal producción deriva de la fundición del mineral. La industria ha utilizado ampliamente el plomo durante décadas en la elaboración de pinturas, tuberías, etc., provocando el aumento de las concentraciones de este metal en el ambiente. Los pingüinos se encuentran en una posición alta dentro de la cadena trófica y, en consecuencia, bioacumulan concentraciones de metales pesados de la cual una parte es depositada en el ambiente a través de restos orgánicos tales como las heces. Con el objetivo de determinar la concentración de plomo en suelos de colonias del Pingüino de Adelia (Pygoscelis adeliae), Pingüino antártico (P. antarctica) y Pingüino papúa (P. papua) en la Península Ardley, se recolectó un total de 27 muestras de suelos en lugares sin pingüinos, lugares de tránsito de pingüinos y en colonias de pingüinos; posteriormente se realizó un análisis de espectrofotometría de absorción atómica. La concentración de plomo, en suelos de zona de tránsito, fue significativamente mayor (p=0,0053) (Media: 4,45 mg/kg) que la concentración de plomo en la zona de ausencia de pingüinos (Media: 3,63 mg/kg). Contrario a lo esperado, la concentración de plomo en suelos de colonias de pingüinos fue menor a suelos de tránsito de pingüinos y no se diferenció significativamente de las zonas de paso (Media: 5,92) y ausencia de pingüinos (p>0,05). La presencia de pingüinos en la Antártica aumenta la capacidad de absorción de plomo en suelos antárticos proveniente de deposición atmosférica.Facultad de Ciencias Naturales y Oceanográfica

Noncoding RNAs: Emerging Players in Muscular Dystrophies

The fascinating world of noncoding RNAs has recently come to light, thanks to the development of powerful sequencing technologies, revealing a variety of RNA molecules playing important regulatory functions in most, if not all, cellular processes. Many noncoding RNAs have been implicated in regulatory networks that are determinant for skeletal muscle differentiation and disease. In this review, we outline the noncoding RNAs involved in physiological mechanisms of myogenesis and those that appear dysregulated in muscle dystrophies, also discussing their potential use as disease biomarkers and therapeutic targets