BASP1 labels neural stem cells in the neurogenic niches of mammalian brain

The mechanisms responsible for determining neural stem cell fate are numerous and complex. To begin to identify the specific components involved in these processes, we generated several mouse neural stem cell (NSC) antibodies against cultured mouse embryonic neurospheres. Our immunohistochemical data showed that the NSC-6 antibody recognized NSCs in the developing and postnatal murine brains as well as in human brain organoids. Mass spectrometry revealed the identity of the NSC-6 epitope as brain abundant, membrane-attached signal protein 1 (BASP1), a signaling protein that plays a key role in neurite outgrowth and plasticity. Western blot analysis using the NSC-6 antibody demonstrated multiple BASP1 isoforms with varying degrees of expression and correlating with distinct developmental stages. Herein, we describe the expression of BASP1 in NSCs in the developing and postnatal mammalian brains and human brain organoids, and demonstrate that the NSC-6 antibody may be a useful marker of these cells.We are grateful to Grigori Enikolopov for critically reviewing the manuscript, Dwight Martin for expert technical assistance, and Huda Zoghbi for the use of flow cytometer. This work was supported by the NIGMS (5R01GM120033), U.S. Army Medical Research (DAMD170110754), Cynthia and Antony Petrello Endowment, and Mark A. Wallace Endowment (M.M.S.); the National Institute of Diabetes and Digestive and Kidney Diseases (T32DK07521-16) (L.N.M.); MINECO SAF-2015-70866R (J.M.E), FPI MICINN predoctoral Fellowship (I.D.); the Proteomics Center at Stony Brook University (NIH/NCRR 1S10 RR023680), and the BCM IDDRC Grant (P50HD10355) from the Eunice Kennedy Shriver National Institute of Child Health and Human Development for use of the Microscopy Core facilities, the RNA In Situ Hybridization Core facility, and the Human Neuronal Differentiation Core facilit

DIAPH3 predicts survival of patients with MGMT-methylated glioblastoma

BackgroundGlioblastoma is one of the most aggressive primary brain tumors, with a poor outcome despite multimodal treatment. Methylation of the MGMT promoter, which predicts the response to temozolomide, is a well-established prognostic marker for glioblastoma. However, a difference in survival can still be detected within the MGMT methylated group, with some patients exhibiting a shorter survival than others, emphasizing the need for additional predictive factors.MethodsWe analyzed DIAPH3 expression in glioblastoma samples from the cancer genome atlas (TCGA). We also retrospectively analyzed one hundred seventeen histological glioblastomas from patients operated on at Saint-Luc University Hospital between May 2013 and August 2019. We analyzed the DIAPH3 expression, explored the relationship between mRNA levels and Patient’s survival after the surgical resection. Finally, we assessed the methylation pattern of the DIAPH3 promoter using a targeted deep bisulfite sequencing approach.ResultsWe found that 36% and 1% of the TCGA glioblastoma samples exhibit copy number alterations and mutations in DIAPH3, respectively. We scrutinized the expression of DIAPH3 at single cell level and detected an overlap with MKI67 expression in glioblastoma proliferating cells, including neural progenitor-like, oligodendrocyte progenitor-like and astrocyte-like states. We quantitatively analyzed DIAPH3 expression in our cohort and uncovered a positive correlation between DIAPH3 mRNA level and patient’s survival. The effect of DIAPH3 was prominent in MGMT-methylated glioblastoma. Finally, we report that the expression of DIAPH3 is at least partially regulated by the methylation of three CpG sites in the promoter region.ConclusionWe propose that combining the DIAPH3 expression with MGMT methylation could offer a better prediction of survival and more adapted postsurgical treatment for patients with MGMT-methylated glioblastoma

Traumatic Brain Injury-induced alterations in Adult Hippocampal Neurogenesis

246 p.Several important cognitive functions affected by traumatic brain injury (TBI) depend on the hippocampus, which harnesses several forms of neural plasticity, among them adult neurogenesis, the generation of new neurons throughout life. Adult hippocampal neurogenesis is a process involved in memory, learning and control of anxiety, cognitive functions which result impaired after TBI. We hypothesize that TBI induces fast and long-term changes in both neural stem cells (NSCs) and newborn neurons which could subsequently alter hippocampal and brain functioning. Using a model of controlled cortical impact (CCI) we have found that TBI has a dual effect on neurogenesis: In the short term (up to two months) it causes an increase in the number of newborn neurons but with aberrant migration, increased soma size and altered electrophysiological properties; in the long term, neurogenesis resultsimpaired by a reduction in the number of immature neurons. We also suggest that the alteration in the expression of Rho Family GTPase 2 (Rnd2) could be causing some of the morphological changes in the immature neurons as well as their aberrant migration and thus could be a target to prevent TBI-induced aberrant neurogenesis, a hypothesis that we are currently investigating at the cellular level. In addition, we have found that NSCs get activated in higher numbers early after TBI, a result that could explain the later reduction in neurogenesis

Presión cricoidea o Sellick, ¿pasado o futuro de una maniobra cuestionada poco evaluada?

Cricoid pressure is an extended maneuver during a rapid sequence induction at certain orotracheal intubations. However, few studies have compared its realization against not doing it. Therefore, there was a need to verify its usefulness or effectiveness. With the present study, which is analyzed with this text, it is not possible to demonstrate the non-inferiority of non-realization. So, a new challenge is achieved in the future: stop doing what we have learned. At least, it will probably be the generations of current residents who will leave as a past practice.La presión cricoidea es una maniobra ampliamente recomendada y extendida a la hora de ciertas intubaciones orotraqueales. Pero son pocos los estudios que hayan comparado su realización frente a no realizarla. Por tanto, aquellos profesionales dedicados a la vía aérea deben comprobar su eficacia. El estudio aquí analizado no consigue demostrar la no inferioridad de la intubación sin presión cricoidea durante la inducción de secuencia rápida (ISR). De modo que se alcanza un nuevo reto en el futuro: dejar de hacer lo que hemos aprendido haciendo. Por lo que probablemente serán las generaciones de residentes actuales los que dejarán como anecdótico una práctica cuestionada pero muy realizada

Image_2_DIAPH3 predicts survival of patients with MGMT-methylated glioblastoma.tif

BackgroundGlioblastoma is one of the most aggressive primary brain tumors, with a poor outcome despite multimodal treatment. Methylation of the MGMT promoter, which predicts the response to temozolomide, is a well-established prognostic marker for glioblastoma. However, a difference in survival can still be detected within the MGMT methylated group, with some patients exhibiting a shorter survival than others, emphasizing the need for additional predictive factors.MethodsWe analyzed DIAPH3 expression in glioblastoma samples from the cancer genome atlas (TCGA). We also retrospectively analyzed one hundred seventeen histological glioblastomas from patients operated on at Saint-Luc University Hospital between May 2013 and August 2019. We analyzed the DIAPH3 expression, explored the relationship between mRNA levels and Patient’s survival after the surgical resection. Finally, we assessed the methylation pattern of the DIAPH3 promoter using a targeted deep bisulfite sequencing approach.ResultsWe found that 36% and 1% of the TCGA glioblastoma samples exhibit copy number alterations and mutations in DIAPH3, respectively. We scrutinized the expression of DIAPH3 at single cell level and detected an overlap with MKI67 expression in glioblastoma proliferating cells, including neural progenitor-like, oligodendrocyte progenitor-like and astrocyte-like states. We quantitatively analyzed DIAPH3 expression in our cohort and uncovered a positive correlation between DIAPH3 mRNA level and patient’s survival. The effect of DIAPH3 was prominent in MGMT-methylated glioblastoma. Finally, we report that the expression of DIAPH3 is at least partially regulated by the methylation of three CpG sites in the promoter region.ConclusionWe propose that combining the DIAPH3 expression with MGMT methylation could offer a better prediction of survival and more adapted postsurgical treatment for patients with MGMT-methylated glioblastoma.</p