Ice-Age Climate Adaptations Trap the Alpine Marmot in a State of Low Genetic Diversity.

Some species responded successfully to prehistoric changes in climate [1, 2], while others failed to adapt and became extinct [3]. The factors that determine successful climate adaptation remain poorly understood. We constructed a reference genome and studied physiological adaptations in the Alpine marmot (Marmota marmota), a large ground-dwelling squirrel exquisitely adapted to the "ice-age" climate of the Pleistocene steppe [4, 5]. Since the disappearance of this habitat, the rodent persists in large numbers in the high-altitude Alpine meadow [6, 7]. Genome and metabolome showed evidence of adaptation consistent with cold climate, affecting white adipose tissue. Conversely, however, we found that the Alpine marmot has levels of genetic variation that are among the lowest for mammals, such that deleterious mutations are less effectively purged. Our data rule out typical explanations for low diversity, such as high levels of consanguineous mating, or a very recent bottleneck. Instead, ancient demographic reconstruction revealed that genetic diversity was lost during the climate shifts of the Pleistocene and has not recovered, despite the current high population size. We attribute this slow recovery to the marmot's adaptive life history. The case of the Alpine marmot reveals a complicated relationship between climatic changes, genetic diversity, and conservation status. It shows that species of extremely low genetic diversity can be very successful and persist over thousands of years, but also that climate-adapted life history can trap a species in a persistent state of low genetic diversity.This work was supported by the Francis Crick Institute which receives its core funding from Cancer Research UK (FC001134), the UK Medical Research Council (FC001134), and the Wellcome Trust (FC001134). CB and AC are supported by the Agence Nationale de la Recherche (project ANR-13-JSV7-0005) and the Centre National de la Recherche Scientifique (CNRS), CB is supported by the Rhône-Alpes region (Grant 15.005146.01). LD is supported by Agence Nationale de la Recherche (project ANR-12-ADAP-0009). TIG is supported by a Leverhulme Early Career Fellowship (Grant ECF-2015-453) and a NERC grant (NE/N013832/1). JMG is supported by a Hertha Finberg Fellowship (FWF T703). LDR is supported by the Diabetes UK RD Lawrence Fellowship (16/0005382)

Molekulare und funktionale Charakterisierung des Galectin-3-bindenden Proteins LGALS3BP in Krebs und der Zentrosomenbiologie

The centrosome acts as the main microtubule-organizing centre in eukaryotic cells. Besides its function in microtubule nucleation it is further implicated in integrating cellular signalling pathways that are essential for cell cycle progression or the response to DNA damage, for instance. In cancer, numerical and structural centrosome aberrations are frequently detected where they contribute to chromosomal instability. Seeking for factors that are functionally relevant for these centrosomal abnormalities, our group established a protein-protein interaction network among centrosomal and cell cycle-regulatory proteins using TAP-MS analysis. Thereby, LGALS3BP, which is deregulated in cancer, was revealed to interact with centrosomal components. The subcellular localization of LGALS3BP to centrosomes and in particular to the proximal part of centrioles was validated by several approaches in this study. LGALS3BP has been previously reported as a secreted glycoprotein with extracellular functions. The present study, however, provides striking evidence that LGALS3BP is involved in the maintenance of centrosome integrity: LGALS3BP upregulation caused centrosome hypertrophy, while depletion led to an accumulation of defective centriolar structures. Supportingly, LGALS3BP deregulation correlated with centrosome abnormalities in certain prostate as well as breast cancer cell lines. Importantly, two rescue experiments revealed a reversion of these phenotypes towards control cells upon restoration of normal LGALS3BP levels. Furthermore, the presence of both LGALS3BP and its interaction partner C-Nap1 was found to be crucial for cell proliferation and viability as simultaneous loss of these proteins caused aneuploidy and apoptosis. An in vitro kinase assay on LGALS3BP peptides revealed AKT1, CHK1 and CHK2 as candidate kinases, which suggests an implication of LGALS3BP in AKT signalling downstream events and DNA damage response. Together, these findings asked for mechanisms that regulate LGALS3BP expression and reasons for its deregulation in cancer. The present study revealed BRD4 as a positive regulator of LGALS3BP gene expression and as a mediator of INFγ-induced LGALS3BP upregulation. Additionally, EZH2 was found responsible for silencing LGALS3BP gene expression in prostate cancer as EZH2 depletion in a respective prostate cancer cell line led to re-expression of LGALS3BP. BRD4 and EZH2 are deregulated in various cancer types, which might explain LGALS3BP deregulation and centrosome aberrations. This study represents the first characterization of intracellular LGALS3BP maintaining centrosome integrity and function. The new insights into LGALS3BP transcriptional regulation now provide opportunities to study if epigenetic therapies targeting LGALS3BP could counteract centrosome aberration in cancer.Das Zentrosom stellt das wesentliche Mikrotubuli-organisierende Zentrum in eukaryotischen Zellen dar. Neben seiner Funktion bei der Mikrotubulinukleation ist es beteiligt an der Integration von zellulären Signaltransduktionswegen, welche beispielsweise für das Fortschreiten des Zellzyklus sowie für die Antwort auf DNS-Schäden wichtig sind. Numerische und strukturelle Aberrationen des Zentrosoms treten sehr häufig während der Krebsentstehung auf und tragen zur Ausbildung von chromosomaler Instabilität bei. Um funktionell relevante Faktoren für diese zentrosomalen Anomalien zu identifizieren, wurde von unserer Arbeitsgruppe ein Proteininteraktionsnetzwerk mittels TAP-MS-Analyse etabliert, welches sich um zentrosomale und Zellzyklus-regulierende Proteine aufbaut. Hierbei wurde LGALS3BP, ein in Krebs dereguliertes Protein, entdeckt, welches mit anderen zentrosomalen Proteinen interagierte. Es konnte durch mehrere Methoden gezeigt werden, dass LGALS3BP am Zentrosom lokalisiert und zwar insbesondere am proximalen Teil der Zentriolen. Zuvor wurde LGALS3BP als ein sekretiertes Glykoprotein mit extrazellulären Funktionen beschrieben. Diese Studie jedoch belegt, dass LGALS3BP auch an der Aufrechterhaltung zentrosomaler Integrität beteiligt ist. Eine Hochregulation von LGALS3BP führte zu zentrosomaler Hypertrophie, wohingegen eine Verminderung von LGALS3BP eine Akkumulation fehlerhafter zentriolärer Strukturen bewirkte. Diese Ergebnisse korrelieren positiv mit LGALS3BP-Fehlexpression und zentrosomalen Aberrationen in verschiedenen Prostata- und Brustkrebszelllinien. Zudem konnte durch die Wiederherstellung normaler LGALS3BP-Expression in zwei Experimenten der jeweilige zentrosomale Phänotyp rückgängig gemacht werden. Es wurde außerdem gezeigt, dass LGALS3BP zusammen mit seinem Interaktionspartner C-Nap1 für die Zellproliferation und -lebensfähigkeit benötigt wird, da das Herunterregulieren beider Proteine zu Aneuploidie und Apoptose führte. Des Weiteren wurden die Kinasen AKT1, CHK1 und CHK2 über eine in vitro Phosphorylierungs-Analyse an LGALS3BP-Peptiden identifiziert, was darauf hindeutet, dass LGALS3BP in AKT-Signalwegen und in Antwortmechanismen auf DNS-Schäden involviert sein könnte. Aufgrund dieser Ergebnisse stellte sich die Frage nach Möglichkeiten der transkriptionellen Regulation von LGALS3BP und nach den Ursachen dessen Deregulierung in Krebs. In dieser Studie VIIZusammenfassung wurde BRD4 als ein positiver Regulator der LGALS3BP-Genexpression ermittelt, welcher auch als Mediator der INFγ-Antwort von LGALS3BP agierte. Zudem wurde EZH2 für die LGALS3BP-Genstilllegung in Prostatakrebs verantwortlich gefunden, da eine Verminderung von EZH2 zur Reexpression von LGALS3BP in einer entsprechenden Prostatakrebszelllinie führte. In verschiedenen Krebsarten werden BRD4 und EZH2 häufig fehlexprimiert, was auch die LGALS3BP-Deregulation und somit die zentrosomalen Anomalien erklären könnte. Die vorliegende Arbeit stellt die erste Charakterisierung von intrazellulärem LGALS3BP dar, welches zentrosomale Integrität und Funktion aufrechterhält. Die neuen Einblicke in die transkriptionelle Regulation von LGALS3BP erlauben es nun experimentell zu ergründen, ob auf LGALS3BP gerichtete epigenetische Therapien Auswirkungen auf die Krebsentwicklung haben und die möglicherweise ursächlichen zentrosomalen Anomalitäten reduzieren

The Cancer Immunotherapy Biomarker Testing Landscape.

CONTEXT.—: Cancer immunotherapy provides unprecedented rates of durable clinical benefit to late-stage cancer patients across many tumor types, but there remains a critical need for biomarkers to accurately predict clinical response. Although some cancer immunotherapy tests are associated with approved therapies and considered validated, other biomarkers are still emerging and at various states of clinical and translational exploration. OBJECTIVE.—: To provide pathologists with a current and practical update on the evolving field of cancer immunotherapy testing. The scientific background, clinical data, and testing methodology for the following cancer immunotherapy biomarkers are reviewed: programmed death ligand-1 (PD-L1), mismatch repair, microsatellite instability, tumor mutational burden, polymerase δ and ε mutations, cancer neoantigens, tumor-infiltrating lymphocytes, transcriptional signatures of immune responsiveness, cancer immunotherapy resistance biomarkers, and the microbiome. DATA SOURCES.—: Selected scientific publications and clinical trial data representing the current field of cancer immunotherapy. CONCLUSIONS.—: The cancer immunotherapy field, including the use of biomarker testing to predict patient response, is still in evolution. PD-L1, mismatch repair, and microsatellite instability testing are helping to guide the use of US Food and Drug Administration-approved therapies, but there remains a need for better predictors of response and resistance. Several categories of tumor and patient characteristics underlying immune responsiveness are emerging and may represent the next generation of cancer immunotherapy predictive biomarkers. Pathologists have important roles and responsibilities as the field of cancer immunotherapy continues to develop, including leadership of translational studies, exploration of novel biomarkers, and the accurate and timely implementation of newly approved and validated companion diagnostics

Hipovitaminose A e anemia ferropriva em gestantes de duas comunidades do Vale do Ribeira (Estado de São Paulo, Brasil) Vitamin A deficiency and iron deficiency anemia in pregnant women of two communities of the Ribeira River Valley (State of S. Paulo, Brazil)

Com o objetivo de estudar deficiências de vitamina A e de ferro em mulheres grávidas, a nível de comunidade, estudou-se 60 gestantes, residentes na zona urbana de Apiaí e Ribeira (Estado de São Paulo), através de inquérito bioquímico, realizando-se dosagens séricas de beta-caroteno, vitamina A, hemoglobina, ferro sérico e capacidade de ligação de ferro, bem como determinação do hematócrito. Os resultados foram comentados e apontaram não haver hipovitaminose A nesta amostra, apesar de haver grande proporção de resultados de caroteno sérico considerados não-normais. Houve maior prevalência de anemia entre gestantes de Ribeira. A comparação dos resultados obtidos entre as gestantes com respectivos grupos-testemunha, apontou freqüência maior de casos considerados não-normais entre as primeiras, em todos os coeficientes levantados.<br>The present study was carried out on sixty pregnant women residing in the urban area of the cities of Apiaí and Ribeira, State of S. Paulo. Samples were taken from each subject in order to study blood levels of beta-carotene, vitamin A, hemoglobin, serum iron and iron binding capacity as well as packed cell volume, with the objective of studying vitamin A and iron deficiency. The results showed that vitamin A deficiency was absent although a large proportion of carotene levels was abnormal. Regarding anemia, prevalence was higher in Ribeira residents. Refering all results to a control group, it was found that frequency was higher in the sample subjects

Preanalytics and Precision Pathology: Pathology Practices to Ensure Molecular Integrity of Cancer Patient Biospecimens for Precision Medicine.

Biospecimens acquired during routine medical practice are the primary sources of molecular information about patients and their diseases that underlies precision medicine and translational research. In cancer care, molecular analysis of biospecimens is especially common because it often determines treatment choices and may be used to monitor therapy in real time. However, patient specimens are collected, handled, and processed according to routine clinical procedures during which they are subjected to factors that may alter their molecular quality and composition. Such artefactual alteration may skew data from molecular analyses, render analysis data uninterpretable, or even preclude analysis altogether if the integrity of specimen is severely compromised. As a result, patient care and safety may be affected, and medical research dependent on patient samples may be compromised. Despite these issues, there is currently no requirement to control or record preanalytical variables in clinical practice with the single exception of breast cancer tissue handled according to the guideline jointly developed by the American Society of Clinical Oncology and College of American Pathologists (CAP) and enforced through the CAP Laboratory Accreditation Program. Recognizing the importance of molecular data derived from patient specimens, the CAP Personalized Healthcare Committee established the Preanalytics for Precision Medicine Project Team to develop a basic set of evidence-based recommendations for key preanalytics for tissue and blood specimens. If used for biospecimens from patients, these preanalytical recommendations would ensure the fitness of those specimens for molecular analysis and help to assure the quality and reliability of the analysis data