2026-01-31 | Features of anesthesia in a child with pyruvate kinase deficiency in an outpatient dental clinic: case report
INTRODUCTION: Pyruvate kinase deficiency is the most common enzymatic disorder of the glycolytic pathway and a leading cause of hereditary nonspherocytic hemolytic anemia. Anesthetic management in patients with pyruvate kinase deficiency carries a risk of specific complications related to impaired energy metabolism and increased susceptibility of erythrocytes and muscle cells to oxidative stress. OBJECTIVE: To present a clinical case demonstrating the successful use of combined inhalational anesthesia in outpatient pediatric dental practice in a child with pyruvate kinase deficiency. MATERIALS AND METHODS: We describe the anesthetic management and specific considerations in the administration of combined inhalational anesthesia during comprehensive dental treatment in a pediatric patient with confirmed pyruvate kinase deficiency. RESULTS: This approach provided adequate anesthesia throughout the dental procedure without the need for intravenous induction with propofol or administration of muscle relaxants, thus minimizing the risk of oxidative stress and mitochondrial dysfunction. CONCLUSIONS: Anesthetic management in patients with pyruvate kinase deficiency should be individualized, with emphasis on minimizing oxidative injury, maintaining adequate oxygen-carrying capacity, and preventing systemic metabolic complications. Inhalational techniques may offer a safe and effective alternative in selected cases.
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2025-10-16 | Harnessing DNA barcoding to enhance and sustain polyclonality in gene-edited hematopoietic stem cells
A challenge in gene editing for hematopoietic stem and progenitor cells (HSPCs) is achieving efficient editing while preserving long-term engraftment and clonal diversity. Tracking edited clones with high resolution is essential to understand the impact of editing on hematopoiesis. We developed a barcoded AAV6 donor template (BC-AAV) to precisely monitor the fate of edited HSPCs following transplantation. Our findings reveal that, despite initial barcode diversity in vitro, human hematopoiesis generated by edited HSPCs transplanted in immunodeficient mice is driven by a limited number of dominant clones. The engraftment of gene-edited cells follows an oligo/polyclonal pattern, indicating that editing does not alter clonal dynamics in this model. Using BC-AAV, we optimized a gene editing protocol for correcting the PKLR gene, responsible for pyruvate kinase deficiency, a rare disorder that causes severe anemia due to red blood energy imbalance. We implemented key improvements. GMP-grade StemSpan AOF medium and StemRegenin-1 increased clonal diversity while maintaining hematopoietic potential. NHEJ inhibitor AZD-7648, significantly boosted editing efficiency in vitro, and a shorter transduction period enhanced engraftment and clonal balance without compromising editing outcomes. This refined strategy for gene editing in human HSPCs optimizes both efficiency and long-term polyclonal dynamics and has important implications for clinical applications.
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2025-07-04 | CRISPR-Cas Systems in Hemolytic Diseases
Hereditary hematological disorders such as β-thalassemia, pyruvate kinase deficiency, and sickle cell disease may be treated by gene therapy. Hematopoietic stem cells (HSCs) are transduced with a corrected version of the altered gene using lentiviral vectors. To make up for the lack of healthy cells brought on by genetic flaws, lentivirus-corrected HSCs produce more of them. The endogenous regulation of the therapeutic gene and the transgene’s integration into the genome, however, are two significant shortcomings in this strategy. Gene editing allows the altered gene to be repaired with greater precision and more directly using less costly, better understood methods such as CRISPR-Cas9 while leaving the rest of the genome untouched. They have been applied to thalassemia, sickle cell disease, pyruvate kinase deficiency and other hereditary erythroid diseases. Experiments have shown CRISPR-Cas9 can fix hemoglobin deficiencies and turn back on fetal globin chains in bone-marrow cells from adults. The application of gene editing showed faster therapeutic responses with cutting edge treatment strategies for red cell disorders that can provide a basis to cure hemolytic abnormalities, as well as other genetic diseases.
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