Unveiling impaired vascular function and cellular heterogeneity in diabetic donor-derived vascular organoids.

Vascular organoids (VOs), derived from induced pluripotent stem cells (iPSCs), hold promise as in vitro disease models and drug screening platforms. However, their ability to faithfully recapitulate human vascular disease and cellular composition remains unclear. In this study, we demonstrate that VOs derived from iPSCs of donors with diabetes (DB-VOs) exhibit impaired vascular function compared to non-diabetic VOs (ND-VOs). DB-VOs display elevated levels of reactive oxygen species (ROS), heightened mitochondrial content and activity, increased proinflammatory cytokines, and reduced blood perfusion recovery in vivo. Through comprehensive single-cell RNA sequencing, we uncover molecular and functional differences, as well as signaling networks, between vascular cell types and clusters within DB-VOs. Our analysis identifies major vascular cell types (endothelial cells [ECs], pericytes, and vascular smooth muscle cells) within VOs, highlighting the dichotomy between ECs and mural cells. We also demonstrate the potential need for additional inductions using organ-specific differentiation factors to promote organ-specific identity in VOs. Furthermore, we observe basal heterogeneity within VOs and significant differences between DB-VOs and ND-VOs. Notably, we identify a subpopulation of ECs specific to DB-VOs, showing overrepresentation in the ROS pathway and underrepresentation in the angiogenesis hallmark, indicating signs of aberrant angiogenesis in diabetes. Our findings underscore the potential of VOs for modeling diabetic vasculopathy, emphasize the importance of investigating cellular heterogeneity within VOs for disease modeling and drug discovery, and provide evidence of GAP43 (neuromodulin) expression in ECs, particularly in DB-VOs, with implications for vascular development and disease.

Investigating the Skill Development of Medical Students in Focused Assessment With Sonography for Trauma (FAST) Ultrasound: A Comparative Analysis Across Different Stages of Medical Training.

INTRODUCTION: Focused assessment with sonography for trauma (FAST) ultrasound (US) is a valuable medical examination used in trauma settings, particularly for rapid responses to events such as natural disasters. Although the efficacy and benefits of FAST in patient care have been extensively studied, there is limited research on training medical students in FAST. Previous studies have found that medical students can proficiently perform a FAST US after two days of training. However, these studies exclusively included first-year medical students without considering variations in their medical knowledge. Particularly, the advantage of medical students having US experience before undergoing FAST training has not been previously examined. OBJECTIVES: Assess the performance and knowledge acquisition of medical students with and without prior US experience after completing a FAST training course. METHODS: The study included a total of 71 students, consisting of 33 males and 38 females, who were between the ages of 18 and 31, with an average age of 24.6 and a standard deviation of 2.4. The inclusion criteria targeted first- and second-year medical school students who participated on a volunteer basis. Students were divided into two groups: group A, consisting of those without prior US experience, and group B, made up of those who had previous US experience. All students completed a pre-training survey to share their comfort and confidence in US use and knowledge. A baseline FAST exam was conducted to establish initial performance. A comprehensive three-hour training session was then provided. Post-training, students performed another FAST exam to assess improvement, followed by a post-training survey to evaluate comfort and confidence. RESULTS: Medical students who had prior experience in the US (group B) performed significantly better (p<0.01) in both the pre- and post-training FAST exams when compared to students without previous US experience. Specifically, in locating the liver, right kidney, hepatorenal recess, and left kidney, as well as detecting fluid accumulation when in a supine position. Additionally, medical students with prior US experience (group B) exhibited higher baseline confidence (p<0.005-p<0.01) in their ability to perform a FAST exam, as indicated by the results of the pre-testing survey. CONCLUSION: Previous experience with US significantly boosted confidence and knowledge gains following FAST training. This emphasizes the value of including US training in medical school programs after earlier exposure, offering evident benefits. The study reveals the unexplored benefit of having prior US experience for medical students undergoing FAST training, thus addressing a previously unexplored area in current research. The conclusions stress the necessity of integrating US training into medical school curricula after initial exposure. This understanding can direct medical educators in refining the education process, enabling students to be better equipped for real-world medical situations involving FAST.

Effects of non-coding RNAs and RNA-binding proteins on mitochondrial dysfunction in diabetic cardiomyopathy.

Vascular complications are the main cause of diabetes mellitus-associated morbidity and mortality. Oxidative stress and metabolic dysfunction underly injury to the vascular endothelium and myocardium, resulting in diabetic angiopathy and cardiomyopathy. Mitochondrial dysfunction has been shown to play an important role in cardiomyopathic disruptions of key cellular functions, including energy metabolism and oxidative balance. Both non-coding RNAs and RNA-binding proteins are implicated in diabetic cardiomyopathy, however, their impact on mitochondrial dysfunction in the context of this disease is largely unknown. Elucidating the effects of non-coding RNAs and RNA-binding proteins on mitochondrial pathways in diabetic cardiomyopathy would allow further insights into the pathophysiological mechanisms underlying diabetic vascular complications and could facilitate the development of new therapeutic strategies. Stem cell-based models can facilitate the study of non-coding RNAs and RNA-binding proteins and their unique characteristics make them a promising tool to improve our understanding of mitochondrial dysfunction and vascular complications in diabetes.

Diabetic endotheliopathy: RNA-binding proteins as new therapeutic targets.

Diabetic Endotheliopathy is widely regarded as a principal contributor to cardiovascular disease pathogenesis in individuals with Diabetes mellitus. The endothelium, the innermost lining of blood vessels, consists of an extensive monolayer of endothelial cells. Previously regarded as an interface, the endothelium is now accepted as an organ system with critical roles in vascular health; its dysfunction therefore is detrimental. Endothelial dysfunction induces blood vessel damage resulting in a restriction of blood and oxygen supply to tissues, the central pathology of cardiovascular disease. Hyperglycemic conditions have repeatedly been isolated as a pivotal inducer of endothelial cell dysfunction. Numerous studies have since proven hyperglycemic conditions to significantly alter the gene expression profile of endothelial cells, with this being largely attributable to the post-transcriptional regulation of RNA-binding proteins. In particular, the RBP Quaking-7 has recently emerged as a crucial mediator of diabetic endotheliopathy, with great potential to become a therapeutic target.

Author Correction: Targeting QKI-7 in vivo restores endothelial cell function in diabetes.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Targeting QKI-7 in vivo restores endothelial cell function in diabetes.

Vascular endothelial cell (EC) dysfunction plays a key role in diabetic complications. This study discovers significant upregulation of Quaking-7 (QKI-7) in iPS cell-derived ECs when exposed to hyperglycemia, and in human iPS-ECs from diabetic patients. QKI-7 is also highly expressed in human coronary arterial ECs from diabetic donors, and on blood vessels from diabetic critical limb ischemia patients undergoing a lower-limb amputation. QKI-7 expression is tightly controlled by RNA splicing factors CUG-BP and hnRNPM through direct binding. QKI-7 upregulation is correlated with disrupted cell barrier, compromised angiogenesis and enhanced monocyte adhesion. RNA immunoprecipitation (RIP) and mRNA-decay assays reveal that QKI-7 binds and promotes mRNA degradation of downstream targets CD144, Neuroligin 1 (NLGN1), and TNF-α-stimulated gene/protein 6 (TSG-6). When hindlimb ischemia is induced in diabetic mice and QKI-7 is knocked-down in vivo in ECs, reperfusion and blood flow recovery are markedly promoted. Manipulation of QKI-7 represents a promising strategy for the treatment of diabetic vascular complications.