Single-nucleus sequencing reveals enriched expression of genetic risk factors in extratelencephalic neurons sensitive to degeneration in ALS.

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder characterized by a progressive loss of motor function linked to degenerating extratelencephalic neurons/Betz cells (ETNs). The reasons why these neurons are selectively affected remain unclear. Here, to understand the unique molecular properties that may sensitize ETNs to ALS, we performed RNA sequencing of 79,169 single nuclei from cortices of patients and controls. In both patients and unaffected individuals, we found significantly higher expression of ALS risk genes in THY1+ ETNs, regardless of diagnosis. In patients, this was accompanied by the induction of genes involved in protein homeostasis and stress responses that were significantly induced in a wide collection of ETNs. Examination of oligodendroglial and microglial nuclei revealed patient-specific downregulation of myelinating genes in oligodendrocytes and upregulation of an endolysosomal reactive state in microglia. Our findings suggest that selective vulnerability of extratelencephalic neurons is partly connected to their intrinsic molecular properties sensitizing them to genetics and mechanisms of degeneration.

Myeloid and lymphoid expression of C9orf72 regulates IL-17A signaling in mice.

A mutation in C9ORF72 is the most common cause of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Patients with ALS or FTD often develop autoimmunity and inflammation that precedes or coincides with the onset of neurological symptoms, but the underlying mechanisms are poorly understood. Here, we knocked out murine C9orf72 in seven hematopoietic progenitor compartments by conditional mutagenesis and found that myeloid lineage C9orf72 prevents splenomegaly, loss of tolerance, and premature mortality. Furthermore, we demonstrated that C9orf72 plays a role in lymphoid cells to prevent interleukin-17A (IL-17A) production and neutrophilia. Mass cytometry identified early and sustained elevation of the costimulatory molecule CD80 expressed on C9orf72-deficient mouse macrophages, monocytes, and microglia. Enrichment of CD80 was similarly observed in human spinal cord microglia from patients with C9ORF72-mediated ALS compared with non-ALS controls. Single-cell RNA sequencing of murine spinal cord, brain cortex, and spleen demonstrated coordinated induction of gene modules related to antigen processing and presentation and antiviral immunity in C9orf72-deficient endothelial cells, microglia, and macrophages. Mechanistically, C9ORF72 repressed the trafficking of CD80 to the cell surface in response to Toll-like receptor agonists, interferon-γ, and IL-17A. Deletion of Il17a in C9orf72-deficient mice prevented CD80 enrichment in the spinal cord, reduced neutrophilia, and reduced gut T helper type 17 cells. Last, systemic delivery of an IL-17A neutralizing antibody augmented motor performance and suppressed neuroinflammation in C9orf72-deficient mice. Altogether, we show that C9orf72 orchestrates myeloid costimulatory potency and provide support for IL-17A as a therapeutic target for neuroinflammation associated with ALS or FTD.

PIKFYVE inhibition mitigates disease in models of diverse forms of ALS.

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease that results from many diverse genetic causes. Although therapeutics specifically targeting known causal mutations may rescue individual types of ALS, these approaches cannot treat most cases since they have unknown genetic etiology. Thus, there is a pressing need for therapeutic strategies that rescue multiple forms of ALS. Here, we show that pharmacological inhibition of PIKFYVE kinase activates an unconventional protein clearance mechanism involving exocytosis of aggregation-prone proteins. Reducing PIKFYVE activity ameliorates ALS pathology and extends survival of animal models and patient-derived motor neurons representing diverse forms of ALS including C9ORF72, TARDBP, FUS, and sporadic. These findings highlight a potential approach for mitigating ALS pathogenesis that does not require stimulating macroautophagy or the ubiquitin-proteosome system.

Efficient generation of lower induced motor neurons by coupling Ngn2 expression with developmental cues.

Human pluripotent stem cells (hPSCs) are a powerful tool for disease modeling of hard-to-access tissues (such as the brain). Current protocols either direct neuronal differentiation with small molecules or use transcription-factor-mediated programming. In this study, we couple overexpression of transcription factor Neurogenin2 (Ngn2) with small molecule patterning to differentiate hPSCs into lower induced motor neurons (liMoNes/liMNs). This approach induces canonical MN markers including MN-specific Hb9/MNX1 in more than 95% of cells. liMNs resemble bona fide hPSC-derived MN, exhibit spontaneous electrical activity, express synaptic markers, and can contact muscle cells in vitro. Pooled, multiplexed single-cell RNA sequencing on 50 hPSC lines reveals reproducible populations of distinct subtypes of cervical and brachial MNs that resemble their in vivo, embryonic counterparts. Combining small molecule patterning with Ngn2 overexpression facilitates high-yield, reproducible production of disease-relevant MN subtypes, which is fundamental in propelling our knowledge of MN biology and its disruption in disease.

Loss of mouse Stmn2 function causes motor neuropathy.

Amyotrophic lateral sclerosis (ALS) is characterized by motor neuron degeneration accompanied by aberrant accumulation and loss of function of the RNA-binding protein TDP43. Thus far, it remains unresolved to what extent TDP43 loss of function directly contributes to motor system dysfunction. Here, we employed gene editing to find whether the mouse ortholog of the TDP43-regulated gene STMN2 has an important function in maintaining the motor system. Both mosaic founders and homozygous loss-of-function Stmn2 mice exhibited neuromuscular junction denervation and fragmentation, resulting in muscle atrophy and impaired motor behavior, accompanied by an imbalance in neuronal microtubule dynamics in the spinal cord. The introduction of human STMN2 through BAC transgenesis was sufficient to rescue the motor phenotypes observed in Stmn2 mutant mice. Collectively, our results demonstrate that disrupting the ortholog of a single TDP43-regulated RNA is sufficient to cause substantial motor dysfunction, indicating that disruption of TDP43 function is likely a contributor to ALS.

Caracterización de los factores de riesgo modificables para el cáncer colorrectal / Characterization of Modifiable Risk Factors for Colorectal Cancer

RESUMEN Fundamento: el cáncer colorrectal constituye un problema sanitario de gran magnitud debido a su elevada morbimortalidad. En Cuba el cáncer de colon constituye la tercera causa de muerte por cáncer en ambos sexos. Objetivo: describir los factores de riesgo modificables para el cáncer colorrectal de los pacientes mayores de 50 años en el consultorio médico de la familia No. 4 del Policlínico Comunitario Docente José Luis Chaviano Chávez de Cienfuegos. Métodos: se realizó un estudio descriptivo de corte transversal que tuvo como universo 318 pacientes que constituyen la totalidad de la población mayor de 50 años del consultorio médico de la familia No. 4 perteneciente al Policlínico Comunitario Docente José Luis Chaviano Chávez de Cienfuegos. La muestra del estudio fue seleccionada por muestreo probabilístico aleatorio simple y estuvo constituida por 152 pacientes. Se estudiaron las variables: edad, sexo y los hábitos modificables: hábito de fumar, consumo de alcohol, consumo de frutas, verduras y vegetales, realización de actividad física, consumo de carne procesada y consumo de carnes rojas. La recogida de la información se realizó por medio de una encuesta. Resultados: de los 318 pacientes del estudio el hábito de fumar estuvo presente en el 38,1 % con predomino del sexo masculino con el 48,6 %. El consumo de alcohol de riesgo se registró en el 40,7 %. El consumo de frutas, verduras y vegetales fue muy bajo con solo 5,9 %, sin diferencias entre ambos sexos. Predominó el índice de masa corporal de riesgo en el 56,7 %, con ligero predominio en el sexo femenino para un 58,5 %, además existió un alto grado de inactividad física con un 91,5 %. Conclusiones: en los pacientes del estudio predominó el hábito de fumar y el consumo de alcohol en el sexo masculino. El consumo de frutas, verduras y vegetales fue muy bajo sin diferencias entre ambos sexos. Predominó el índice de masa corporal de riesgo con ligera superioridad en el sexo femenino, existió un alto grado de inactividad física.

ABSTRACT Background: colorectal cancer constitutes a great magnitude health problem due to its high morbidity and mortality. In Cuba, colon cancer is the third leading cause of cancer death in both sexes. Objective: to describe the modifiable risk factors for colorectal cancer in patients over 50 years of age in the José Luis Chaviano Chávez Teaching Community Polyclinic Medical office of Family No. 4 in Cienfuegos. Methods: a descriptive, cross-sectional study was carried out with a universe of 318 patients that constitute the entire population over 50 years of the family No. 4 medical office belonging to the José Luis Chaviano Chávez Polyclinic in Cienfuegos. The study sample was selected by simple random probability sampling, and consisted of 152 patients. The studied variables: age, sex, and modifiable habits: smoking, alcohol consumption, consumption of fruits, vegetables and vegetables, physical activity, consumption of processed meat and consumption of red meat. The information was collected through a survey. Results: of the 318 patients in the study, smoking was present in the 38.1 %, predominantly male with the 48.6 %. Risk alcohol consumption was recorded in the 40.7 %. The consumption of fruits, vegetables and vegetables was very low with only 5.9 %, without differences between both sexes. The risk body mass index predominated in 56.7 %, with a slight predominance in the female sex for 58.5 %; there is also a high degree of physical inactivity with 91.5 %. Conclusions: male smoking and alcohol consumption predominated in the study patients. The consumption of fruits and vegetables was very low without differences between both sexes. The risk body mass index predominated with a slight superiority in the female sex; there was a high degree of physical inactivity.

C9orf72 suppresses systemic and neural inflammation induced by gut bacteria.

A hexanucleotide-repeat expansion in C9ORF72 is the most common genetic variant that contributes to amyotrophic lateral sclerosis and frontotemporal dementia1,2. The C9ORF72 mutation acts through gain- and loss-of-function mechanisms to induce pathways that are implicated in neural degeneration3-9. The expansion is transcribed into a long repetitive RNA, which negatively sequesters RNA-binding proteins5 before its non-canonical translation into neural-toxic dipeptide proteins3,4. The failure of RNA polymerase to read through the mutation also reduces the abundance of the endogenous C9ORF72 gene product, which functions in endolysosomal pathways and suppresses systemic and neural inflammation6-9. Notably, the effects of the repeat expansion act with incomplete penetrance in families with a high prevalence of amyotrophic lateral sclerosis or frontotemporal dementia, indicating that either genetic or environmental factors modify the risk of disease for each individual. Identifying disease modifiers is of considerable translational interest, as it could suggest strategies to diminish the risk of developing amyotrophic lateral sclerosis or frontotemporal dementia, or to slow progression. Here we report that an environment with reduced abundance of immune-stimulating bacteria10,11 protects C9orf72-mutant mice from premature mortality and significantly ameliorates their underlying systemic inflammation and autoimmunity. Consistent with C9orf72 functioning to prevent microbiota from inducing a pathological inflammatory response, we found that reducing the microbial burden in mutant mice with broad spectrum antibiotics-as well as transplanting gut microflora from a protective environment-attenuated inflammatory phenotypes, even after their onset. Our studies provide further evidence that the microbial composition of our gut has an important role in brain health and can interact in surprising ways with well-known genetic risk factors for disorders of the nervous system.

Utilización del biomarcador de cistatina C en pacientes conposible fallo renal / Use of Cystatin C Biomarker in Patients with Possible Renal Failure

RESUMEN La lesión renal es una complicación frecuente en los pacientes ambulatorios y hospitalizados y su incidencia ha aumentado en los últimos años, ya sea como una enfermedad primaria o como diagnóstico secundario. La detección precoz de laenfermedad renal en sus grados más leves resultaría beneficiosapara comenzar con tratamiento. La presente revisión bibliográfica, sustentada en el análisis de cerca de 30 artículos científicos, describe la factibilidad del uso de la cistatina C como biomarcador renal; se refiere además a las ventajas y limitaciones que reporta su utilización dentro del bioanálisis clínico; y por último la importancia del uso futuro de la cistatina C como biomarcador analítico en los diferentes laboratorios del sistema de salud enCienfuegos. Este trabajo aporta información relevante acerca de la actualización y preparación del personal de salud en afecciones que pueden ser detectadas a nivel molecular y celular.

ABSTRACT Renal injury is a frequent complication in hospitalized and outpatients and its incidence has increased in recent years, either as a primary disease or as a secondary diagnosis. Early detection of kidney disease in its milder degree would be beneficial to begin treatment for better results. This literature review, based on the analysis of about 30 scientific articles, describes the feasibility of using cystatin C as a renal biomarker; it also refers to the advantages and limitations that its use reports within the clinical bioanalysis; and finally the importance of the future use of cystatin C as an analytical biomarker in different laboratories of the health system in Cienfuegos. This work provides relevant information about health personnel´s updating and training in medical conditions which may be detected at the molecular and cellular level.

Telomerasa: fuente de juventud para la célula / Telomerase and cells youth

La telomerasa es una enzima formada por un complejo proteína - ácido ribonucleico (ARN) que permite el alargamiento de los telómeros, estructuras de ADN no codificantes. Se recopilaron los datos significativos y actuales sobre el estudio de la enzima en el proceso del envejecimiento y cómo influyen los telómeros en el mismo, teniendo en cuenta su activación, tiempo de vida, así como sus propiedades. La telomerasa al activarse en las células, permite su rejuvenecimiento y prolonga más su tiempo de vida y su número posible de divisiones en el organismo, constituyendo estas propiedades de la enzima un recurso importante para la prevención y tratamiento de varias enfermedades. Este estudio constituye una fuente valiosa de información para alcanzar una mejor calidad de vida en nuestra población, mejorando los métodos de prevención y tratamiento de enfermedades como el cáncer(AU)

Telomerase is an enzyme formed by a protein-ribonucleic acid complex that allows telomere enlargement, non codifying DNA structures. Cellular activation allows its renovation and extends its life time and its possible number of divisions within the organism. These properties of the enzyme are an important factor in prevention and treatment of various diseases. Significant and updated data is collected about the study of this enzyme in ageing process, life time and properties(AU)

Prevalencía y características de la hemoglobina AS en individuos que acuden al Banco de Sangre / Prevalence and characteristics of AS hemoglobin in individuals attending the blood bank

Se evaluó la frecuencia de la variante hemoglobina AS en individuos que acudieron al banco de sangre provincial de Cienfuegos, y se determinó la asociación entre esta variante y los casos rechazados. Se estudiaron 900 casos procedentes del municipio de Cienfuegos durante el período enero-marzo de 2005. Se empleó el diseño caso y control en su variante estudio de prevalencia. Fueron “casos” los pacientes con hemoglobina AS y “controles” los de hemoglobina AA. A todos, se les realizó examen e interrogatorio médico y pruebas de laboratorio convencionales, que incluían electroforesis de hemoglobina. La prevalencia del rasgo AS fue del 5,2 por ciento y se demostró que los individuos rechazados o no del fenotipo AS, no constituyeron motivo de exclusión de las donaciones. El rasgo AS predominó entre personas de raza negro-mestiza, con diferencia significativa en cifras bajas de hemoglobina, aunque no resulta un índice de riesgo en la aparición de este rasgo(AU)

The frequency of the AS hemoglobin variant in individuals attending the provincial blood bank in Cienfuegos was assessed, and the relationship between this variant and the rejected cases was determined. A total of 900 cases from Cienfuegos municipality were studied during the period January-March 2005. The case and control design in its variant of study of prevalence was implemented. The patients with AS hemoglobin were the cases and those with AA hemoglobin were the control. All the subjects received a medical examination and were interviewed. They were also submitted to conventional laboratory tests, including hemoglobin electrophoresis. The prevalence of the AS trait was of a 5.2 per cent and it was demonstrated that the individuals of AS phenotype, rejected or not, did not constituted a motive for exclusion in the blood donations. The AS trait predominated among persons of African descent with a significant difference in low level hemoglobin, though it is not a risk index in the appearance of this trait(AU)