Adrenal glands stem cells: general signaling pathways

Nowadays stem cells of adult type are attractive in case of active development of cell and genome technologies. They are the target of new therapeutic approaches, which are based on correction of mutations or replenishment of organs, that were damaged by autoimmune reactions, aging or other pathological processes. Also stem cells, including patient-specific (induced Pluripotent Stem Cells, iPSCs), and obtained by differentiation from them tissue cultures and organoids are the closest models to in vivo researches on humans, which gives an opportunity to get more relevant data while testing different therapeutic approaches and pharmacological drugs. The main molecular pathways, that are essential for homeostasis of a cortex of a adrenal gland — compound, structurally and functionally heterogeneous organ, is described the presented review. The adrenal cortex is renewing during the organism’s ontogenesis at the expense of the pool of stem and progenitors cells, which are in tight junctions with differentiated steroidogenic cells and which are under constant control of endocrine and paracrine signals. The understanding of signaling pathways and interactions of different cell types will give an opportunity to develop the most suitable protocols for obtaining cells of adrenal gland cortex in a different stages of differentiation to use them in scientific and medical purposes.

Neurological disorders in patients with long COVID syndrome and cell therapy methods for their correction a literature review

The review presents the current understanding of the incidence and nature of neurological disorders in patients with the so-called long COVID syndrome. Symptoms, putative pathophysiological mechanisms, risk factors, search for methods of treatment and rehabilitation of patients using the patient's own hematopoietic cells are discussed. A search was carried out for scientific articles, including those published in peer-reviewed journals indexed in PubMed, Web of Science, Scopus and RSCI. The inclusion of stem cells (SC) in rehabilitation programs for patients with various injuries and diseases of the central nervous system (CNS) is a promising area of research. The mechanisms of CNS damage therapy based on the use of adult-type pluripotent stem cells, including CD34+, consist of many aspects. On the background of SC transplantation, damaged nerve cells and surrounding tissues, including neurons and glial cells, can be restored, which helps to ensure the integrity of the nerve conduction pathway and, thus, restore nerve function. SC therapy can suppress genes involved in inflammation and apoptosis, as well as activate genes with neuroprotective action, thereby protecting spinal neurons from secondary damage. This line of cell therapy can be used to treat long COVID syndrome.

MPC-1 DNA methylome analysis suggested the presence of “true” IDH-wildtype lower-grade gliomas

Background: There will be significant changes in the diagnosis of IDH-wildtype adult-type gliomas in the upcoming 5th edition of the WHO Classification of Central Nervous System Tumours. IDH-wildtype lower grade gliomas (IDHwt LGGs) that harbor molecular features of glioblastoma (EGFR amplification, the combination of whole chromosome 7 gain and whole chromosome 10 loss (7+/10-), or TERT promoter mutations) will be diagnosed as glioblastomas (GBMs), while IDH-wildtype astrocytomas will not be included as a separate tumor type. However, IDHwt LGGs are a very heterogeneous group of tumors, and further investigation is warranted particularly in those without molecular features of glioblastoma. To elucidate the biology of IDHwt LGGs, we analyzed DNA methylation profile and survival time. Materials and Methods: Of the 724 adult-type diffuse glioma samples from a multi-institutional study, 64 IDHwt LGG, including 54 without any of molecular features of GBM and 10 with PDGFRA amplification or TERT promoter mutation, were examined using Infinium MethylationEPIC BeadChip. The raw data files (IDAT files) were analyzed by the web-based DNA methylation classifier provided by DKFZ (MolecularNeuropathology.org) or by R (Version 4.0.4) using the minfi (1.34.0) and Rtsne (0.15) packages. [Result] Twenty-three out of 54 IDHwt LGGs matched known methylation classes using the DKFZ methylation classifier. In t-Distributed Stochastic Neighbor Embedding clustering analysis, 20 cases formed a cluster within the methylation class family glioblastoma, IDH-wildtype, mainly subclass RTK I (“GBM” cluster). Another 29 IDHwt LGGs formed an independent cluster (“LGG” cluster) separate from any of the existing reference groups near but not overlapping with several subtypes of pediatric-type lower grade gliomas. The “LGG” cluster cases had significantly longer overall survival than the “GBM” cluster cases. Discussion: Methylation profiling showed that IDHwt LGGs without molecular features of GBM were heterogeneous group of tumors. Our data suggested the presence of “true” IDHwt LGGs with intermediate prognosis.

The association between caregiver education on adult T2DM and patient’s outcomes in community: A systematic review

Introduction: Adult type 2 diabetes (T2DM) threatens public health and most patients manage their diabetic condition while in the community. As it is challenging for patients to properly manage diabetes alone, caregiver involvement in T2DM patient care is encouraged. This study aimed to examine the association between caregiver involvement in T2DM education within a community and the patients’ diabetes care outcomes (e.g., glycated hemoglobin (HbA1c) level, behavior, or hospitalization). Methods: The available scientific literature in PubMed, Cochrane, EMBASE, and CINAHL was searched. The methodological quality of bias was assessed using the Cochrane risk of bias tool. Results: A total of 13 out of 741 published studies were synthesized in this review. There is evidence that caregiver involvement in T2DM education is effective in the reduction of HbA1C and BMI, but not necessarily effective in reducing lipids. Study results indicate that caregiver related interventions can significantly improve patient diabetes knowledge, physical activity, and self-efficacy, but results were more mixed regarding medication adherence. Risk of bias analysis classified the majority of studies (77%) to be moderate or high quality. Conclusion: This review aimed to explore the association between caregiver involvement in adult T2DM education in the community and patients’ diabetes care outcomes. The findings show an improvement in biological and behavioral self-management outcomes with caregivers involved in T2DM education, though no studies examined the direct association between complications or hospital readmission. Future research focused on tailored interventions and longer follow-up of patient outcomes are recommended.

Diagnosis of adult ALCAPA with computed tomography coronary artery

Background Anomalous left coronary artery from the pulmonary artery (ALCAPA) is a rare congenital anomaly where the left main coronary artery arises from the pulmonary artery, instead of the coronary sinus of the ascending aorta. ALCAPA is divided into infant and adult types. Life-threatening complications such as malignant arrhythmia and sudden death could ensue in adult type of ALCAPA. Imaging is the current preferred modality for ALCAPA. It is indeed a challenge to diagnose ALCAPA due to its non-specific clinical presentation and laboratory findings. We report a case of adult type ALCAPA presented with unstable angina surprisingly surviving into adulthood with no symptoms prior to presentation, which is extremely rare. Case presentation A 53-year-old lady presented with sudden onset of chest pain and worsening shortness of breath. She was initially treated as unstable angina. Physical examination revealed normal heart sound. Computed tomography coronary angiography (CTCA) showed an anomalous origin of the left coronary artery from the posterior wall of the proximal pulmonary artery, compatible with ALCAPA. The anomalous left coronary artery bifurcates into left anterior descending and left circumflex arteries. Cardiothoracic surgeon planned for occlusion of ALCAPA via the pulmonary artery, owing to the fact that unfeasible rerouting in the presence of well-established collateral supply. Conclusions ALCAPA is a rare and life-threatening condition in adults which may lead to myocardial infarction and sudden death in untreated cases. CTCA is one of the preferred modern imaging modality in ALCAPA owing to its superior ability for direct visualization of the anomaly. Hence, early identification and surgical intervention of the anomaly are paramount to reduce the morbidity and mortality.

Methylation profiling reveals novel molecular classes of rhabdomyosarcoma

Rhabdomyosarcomas (RMS) represent a family of aggressive soft tissue sarcomas that present in both children and adults. Pathologic risk stratification for RMS has been based on histologic subtype, with poor outcomes observed in alveolar rhabdomyosarcoma (ARMS) and the adult-type pleomorphic rhabdomyosarcoma (PRMS) compared to embryonal rhabdomyosarcoma (ERMS). Genomic sequencing studies have expanded the spectrum of RMS, with several new molecularly defined entities, including fusion-driven spindle cell/sclerosing rhabdomyosarcoma (SC/SRMS) and MYOD1-mutant SC/SRMS. Comprehensive genomic analysis has previously defined the mutational and copy number spectrum for the more common ERMS and ARMS and revealed corresponding methylation signatures. Comparatively, less is known about epigenetic correlates for the rare SC/SRMS or PRMS histologic subtypes. Herein, we present exome and RNA sequencing, copy number analysis, and methylation profiling of the largest cohort of molecularly characterized RMS samples to date. In addition to ARMS and ERMS, we identify two novel methylation subtypes, one having SC/SRMS histology and defined by MYOD1 p. L122R mutations and the other matching adult-type PRMS. Selected tumors from adolescent patients grouped with the PRMS methylation class, expanding the age range of these rare tumors. Limited follow-up data suggest that pediatric tumors with MYOD1-mutations are associated with an aggressive clinical course.

Interrogating Post-Transcriptional Mechanisms of Fetal Hemoglobin Regulation

Elevated levels of fetal hemoglobin (HbF) significantly ameliorate clinical outcomes for patients with beta-hemoglobinopathies, such as sickle cell disease (SCD). The only FDA-approved drug for treating SCD through inducing HbF is hydroxyurea, however the mechanism of action is unknown with variable effectiveness among patients. Thus, there remains a strong interest to identify more robust means of upregulating HbF, such as specific inhibition of HbF repressors. BCL11A and LRF are well-characterized transcription factors that independently repress the fetal type b-globin like genes HBG1 and HBG2 but their therapeutic potential is limited by challenging druggability and critical developmental function. However, upstream regulation of these factors, such as post-transcriptional mechanisms, are not well studied and may house novel therapeutic targets. To this end, we employed a CRISPR/Cas9 based screening approach to interrogate a library of RNA binding proteins (RBP) in the context of HbF regulation. Using HUDEP2 cells, a human adult-type erythroid progenitor cell line, we screened 341 human RBPs and identified four candidate RBPs, none of which have previously been implicated in HbF regulation. Of these candidates, RNA Binding Motif 12 (RBM12) showed the greatest level of HbF induction following in vitro depletion. Depletion of RBM12 protein in HUDEP2 cells and human CD34 + hematopoietic stem and progenitor cells (HSPC) via CRISPR/Cas9 editing raised HbF production 2-4 fold as assessed by HbF flow cytometry, HBG1/2 mRNA, and protein (γ-globin). Cell viability and maturation of RBM12 perturbed cells were largely intact. Additionally, RBM12 depletion in CD34 + HSPCs derived from SCD patients resulted in reduced percentage of sickled cells under hypoxic conditions. Unexpectedly, reduction of RBM12 had minimal effect on BCL11A and LRF expression suggesting that RBM12 may regulate HbF through a pathway that is indirectly related or independent of these transcription factors. RBM12 is an RBP that is widely expressed across diverse cell types and contains multiple RNA recognition motifs (RRM). While it has been implicated in various cancers and neurological disorders, its functions are not well studied. As an RBP, RBM12 can carry out several roles of post-transcriptional regulation, such as pre-mRNA splicing, mRNA transport, stabilization, and translation. As these activities are executed in different cellular compartments, we set out to narrow down RBM12 function by assessing its subcellular localization. Immunofluorescence staining revealed strong nuclear presence of RBM12, suggesting that it functions via mRNA biogenesis and/or processing. RNASeq and LC-MS/MS analysis of RBM12 KO CD34 + HSPCs revealed modest changes in the transcriptome and proteome. In order to gain mechanistic insight into RBM12 in the context of HbF regulation, we performed cDNA rescue experiments in RBM12-deficient HUDEP2 clones. Overexpression of full length RBM12 restored HbF repression. Notably, four out of the five RRMs were dispensable for HbF silencing, but RRM1 was essential for this activity. Interestingly, an extended form of RRM1 was also sufficient for HbF silencing. Mechanistic studies of this RRM1 module are underway and will be discussed. In sum, the identification of RBM12 as a regulator of HbF production represents a previously undescribed post-transcriptional layer of hemoglobin gene regulation. In pursuing this path, we hope to gain a deeper understanding of this understudied RBP in the context of HbF regulation which might in turn lead to the identification of potential therapeutic targets for the treatment of SCD and other hemoglobinopathies. Disclosures Blobel: Pfizer: Consultancy; Fulcrum Therapeutics, Inc.: Consultancy.