Damaging mutations of the Angiotensin I-converting enzyme (ACE) that result in low ACE levels may increase the risk of developing late-onset Alzheimer’s disease (AD). We quantified blood ACE levels in EDTA-plasma from 147 subjects with 23 different heterozygous ACE mutations (and 70 controls) and estimated the effect of these mutations on ACE phenotype, using a set of monoclonal antibodies (mAbs) to ACE and two ACE substrates. We identified several mutations in both ACE domains (including the most frequent ACE mutation, Y215C), which led to decreased ACE levels in the blood, and thus could be considered as putative risk factors for late-onset AD. The precipitation of several ACE mutants (Q259R, A725P, C734Y) by specific mAbs changed significantly, and therefore, these mAbs could be markers of these mutations. Analysis of 50 of the most frequent ACE mutations demonstrates that more than 1.5% of the adult population may have mutations which lead to decreased ACE levels, and thus, the role of low ACE levels in the development of AD may be underappreciated. Intriguingly, statistical and cluster analyses of longevity patients revealed trends towards higher frequency of cognitive impairment among affected individuals with damaging ACE mutations. Systematic analysis of blood ACE levels in patients with various ACE mutations identifies individuals with low blood ACE levels who may be at increased risk for late-onset AD. Patients with transport-deficient ACE mutations theoretically could benefit from therapeutic treatment with a combination of chemical and pharmacological chaperones and proteasome inhibitors, as was demonstrated previously on a cell model of the transport-deficient ACE mutation Q1069R. Moreover, clinical association analysis suggests a trend linking damaging ACE mutations with increased risk of cognitive impairment.

The expanded newborn screening (NBS) program in the Russian Federation, launched in 2023, includes the detection of severe forms of T- and B-cell immunodeficiencies via TREC/KREC quantification. We report a rare case of a male infant having multiple congenital anomalies and lymphopenia identified through this program. Genetic testing revealed a 25.8 Mb terminal deletion spanning 13q31.2–qter, consistent with 13q deletion syndrome. Initial NBS revealed reduced TREC levels, prompting further evaluation. The patient exhibited a complex phenotype, including central nervous system malformation (alobar holoprosencephaly), severe congenital heart disease, renal hypoplasia, limb and genitourinary anomalies, and facial dysmorphism. Postnatal complications included pneumonia, pleuritis, and chylothorax. Flow cytometry demonstrated mild T- and B-cell lymphopenia. The genomic defect was characterized using long-read third-generation sequencing, enabling precise breakpoint identification and accurate mapping of deleted genes. The deletion was confirmed via subtelomeric FISH analysis. The patient died at 7 months of age due to the progression of underlying congenital anomalies and associated complications. Our findings broaden the clinical characterization of distal 13q deletion syndrome and demonstrate the value of long-read sequencing in structural chromosomal analysis. They further highlight the difficulties of caring for neonates having complex malformations and immune dysfunction. Given the potential for both primary and secondary immune disturbances, comprehensive immunological evaluation should be considered in patients having 13q deletion syndrome to improve diagnostic accuracy and inform appropriate clinical management.

Introduction. Bacteria are capable of integrating a portion of the genetic material of an invading infectious agent as a spacer into the CRISPR array. This process is called CRISPR adaptation and is necessary for bacteria to adequately respond to the same infectious agent in the future. The molecular mechanisms of CRISPR adaptation remain unclear, particularly the role of extra-CRISPR-Cas components in bacteria during prespacer generation and spacer acquisition. In this study, we developed an approach to identifying nucleases responsible for the formation of prespacer 3’-ends in vivo. Materials and methods. We generated a genetic construct based on the pBR322 plasmid vector with gRNAs targeting the genes encoding cellular exonucleases with 3’-5’ activity in E. coli. This genetic construct and the pdCas9 plasmid were transformed into KD403 cells. The level of primed CRISPR adaptation was assessed using PCR and sequencing. The structure of the 3' ends of the prespacers was assessed using the FragSeq short-fragment sequencing method using the Accel-NGS® 1S Plus DNA Library Kit, Swift biosciences (xGen™ ssDNA & Low-Input DNA Library Preparation Kit, IDT). Results. In the KD403 ΔrecB strain with suppressed nucleases, a reduction in primed adaptation was detected, but sequencing revealed no disruption of the 3’ ends of the spacer precursors. Conclusion. Our results contribute to the understanding of the molecular mechanisms of CRISPR adaptation, which may aid in the development of CRISPR-based approaches in medicine and agriculture in the future.

Introduction. Bacteria are capable of integrating a portion of the genetic material of an invading infectious agent as a spacer into the CRISPR array. This

Identifying genetic cause(s) is a key step for management and treatment of patients with inborn errors of immunity (IEI). Here, in an observational cross-sectional genomic study, we analyzed whole-genome sequencing (WGS) data of 72 IEI patients from Saint Petersburg and Northwestern Russia: 42 patients with common variable immunodeficiency (CVID)-like phenotypes, 6 patients with clinically diagnosed X-linked agammaglobulinemia (XLA or Bruton’s disease), and 24 patients with other forms of IEI. Causative pathogenic and likely pathogenic variants in BTK, CYBB, CHD7, AIRE, ATM, SBDS, NFKB1, and CTLA4 genes were identified in 14 (19%) patients. Variants of uncertain significance that could be linked to observed clinical phenotypes were detected in 6 patients. These included a BTK variant in a patient with Bruton’s disease, variants in SH2D1A, SOCS1, and IKBKB in patients with CVID, and variants in CARD11 and CD40LG in patients with other forms of IEI. Additional rare variants that were mostly unique to individual patients were found in multiple IEI genes from the International Union of Immunological Societies (IUIS) Expert Committee 2024 list. In the CVID-like subcohort, pathway-level analysis of these rare variants revealed patterns associated with clinical manifestations. Taken together, our results expand the genetic characterization of an understudied regional IEI cohort, particularly of patients with CVID-like phenotypes, and identify genetic factors that are implicated in or may contribute to the disease.

Cryopyrin-associated periodic syndrome is a group of autoinflammatory diseases caused by mutations in the NLRP3 gene, which lead to uncontrolled inflammasome activation and, consequently, excessive production of the proinflammatory cytokine interleukin 1β. Diagnosis is challenging, since many patients have a heterogeneous multisystem clinical picture. This publication presents a familial case of the disease with variability of manifestations from “blurred” and asymptomatic to a detailed clinical picture with multisystem involvement (persistent inflammatory activity, fever and urticaria associated with cold exposure, erythema nodosum, poly- and monoarthritis, uveitis, aphthous stomatitis, scrotal ulcers, and headache). Two patients had an atypical presentation of symptoms with predominant involvement of the joints, eyes, and oral mucosa, due to which one of them was initially given an alternative diagnosis of Behcet’s disease (syndrome). Genetic testing made it possible to identify a pathogenic mutation in the NLRP3 gene, verify the diagnosis, and promptly prescribe pathogenetic therapy with an interleukin 1 inhibitor, which is necessary to prevent irreversible organ damage (amyloidosis, sensorineural hearing loss, vision loss, etc.) and reduce the burden of the disease.