Tubulin, one of the most abundant cytoskeletal building blocks, has numerous isotypes in metazoans encoded by different conserved genes. Whether these distinct isotypes form cell type– and context-specific microtubule structures is poorly understood. Based on a cohort of 12 patients with primary ciliary dyskinesia as well as mouse mutants, we identified and characterized variants in the TUBB4B isotype that specifically perturbed centriole and cilium biogenesis. Distinct TUBB4B variants differentially affected microtubule dynamics and cilia formation in a dominant-negative manner. Structure-function studies revealed that different TUBB4B variants disrupted distinct tubulin interfaces, thereby enabling stratification of patients into three classes of ciliopathic diseases. These findings show that specific tubulin isotypes have distinct and nonredundant subcellular functions and establish a link between tubulinopathies and ciliopathies.

Integrin-mediated adhesion to the extracellular matrix is a key regulator of the cell cycle, as demonstrated for the passage of the G1/S checkpoint and the completion of cytokinetic abscission. Here, integrin-dependent regulation of the cell cycle in G2 and early M phases was investigated. The progression through the G2 and M phases was monitored by live-cell imaging and immunofluorescence staining in adherent and non-adherent fibroblast cells. Non-adherent cells, as well as adherent cells lacking FAK activity due to suppressed expression or pharmacological inhibition, exhibited a prolonged G2 phase and severely defect centrosome separation, resulting in delayed progress through the early mitotic stages. The activation of the critical mitotic regulator PLK1 and its indirect target Eg5, a kinesin-family motor protein driving the centrosome separation, were reduced in the cells lacking FAK activity. Furthermore, the absence of integrin adhesion or FAK activity destabilized the structural integrity of centrosomes and often caused detachment of pericentriolar material from the centrioles. These data identify a novel adhesion-dependent mechanism by which integrins via FAK and PLK1 contribute to the regulation of the cell cycle in the G2 and early M phases, and to the maintenance of genome integrity.

Primary ciliary dyskinesia (PCD) is a genetic condition that results in dysmotile cilia. The repercussions of cilia dysmotility and gene variants on the multiciliated cell remain poorly understood. We used single-cell RNA-Seq, proteomics, and advanced microscopy to compare primary culture epithelial cells from patients with PCD, their heterozygous mothers, and healthy individuals, and we induced pluripotent stem cells (iPScs) generated from a patient with PCD. Transcriptomic analysis revealed unique signatures in PCD airway cells compared with their mothers’ cells and the cells of healthy individuals. Gene expression in heterozygous mothers’ cells diverged from both control and PCD cells, marked by increased inflammatory and cellular stress signatures. Primary and iPS-derived PCD multiciliated cells had increased expression of glutathione-S-transferases GSTA2 and GSTA1 , as well as NRF2 target genes, accompanied by elevated levels of reactive oxygen species (ROS). Immunogold labeling in human cilia and proteomic analysis of the ciliated organism Chlamydomonas reinhardtii demonstrated that GSTA2 localizes to motile cilia. Loss of human GSTA2 and C . reinhardtii GSTA resulted in slowed cilia motility, pointing to local cilia regulatory roles. Our findings identify cellular responses unique to PCD variants and independent of environmental stress and uncover a dedicated ciliary GSTA2 pathway essential for normal motility that may be a therapeutic target.

Chimeric antigen receptor (CAR)–based therapies have pioneered synthetic cellular immunity but remain limited in their long-term efficacy. Emerging data suggest that dysregulated CAR-driven T-cell activation causes T-cell dysfunction and therapeutic failure. To re-engage the precision of the endogenous T-cell response, we designed MHC-independent T-cell receptors (miTCR) by linking antibody variable domains to T-cell receptor constant chains. Using predictive modeling, we observed that this standard “cut and paste” approach to synthetic protein design resulted in myriad biochemical conflicts at the hybrid variable–constant domain interface. Through iterative modeling and sequence modifications, we developed structure-enhanced miTCRs which significantly improved receptor-driven T-cell function across multiple tumor models. We found that 41BB costimulation specifically prolonged miTCR T-cell persistence and enabled improved leukemic control in vivo compared with classic CAR T cells. Collectively, we have identified core features of hybrid receptor structure responsible for regulating function. Significance: Improving the durability of engineered T-cell immunotherapies is critical to enhancing efficacy. We used a structure-informed design to evolve improved miTCR function across several models. This work underscores the central role of synthetic receptor structure in T-cell function and provides a framework for improved receptor engineering.

Alzheimer's disease (AD) is a neurodegenerative disorder characterized by progressive cognitive decline and the accumulation of extracellular amyloid-beta (Aβ) plaques and intracellular neurofibrillary tangles in the brain tissue. Like other dementias, AD disproportionately affects certain racial and ethnic groups, remaining a major global health challenge that demands improved strategies for early detection, mechanistic understanding, and therapeutic intervention. This special issue focuses on studies investigating microglia dysfunction in various neurological disorders, including AD, while considering factors like experimental models, sex, age, and ethnicity. These studies leverage advanced computational methods to integrate and analyze molecular data. Such approaches, including multi-omics analyses (shown in Figure 1), are elucidating AD's molecular landscape and enabling personalized strategies for neurodegenerative diseases, such as developing genetic and molecular biomarkers, identifying novel therapeutic targets, and designing more effective clinical trials. Ultimately, this research is essential for advancing our understanding of microglial function and dysfunction across diverse populations. In this special issue, we included six articles, including two reviews and four research articles, discussed briefly below. Two studies focus on the marker identification, highlighting potential for improved diagnosis and therapeutic targeting in AD. Li Wu et.al demonstrate that bloodderived cell-free RNA (cfRNA) can serve as a non-invasive biomarker for detecting molecular changes associated with AD. By integrating cfRNA profiles with single-cell transcriptomic (scRNA) and machine learning (ML) approaches, the authors identified a panel of 34 signature genes, achieving robust diagnostic performance (~90% AUC). These findings underscore the potential of these signature genes for early, non-invasive AD detection and patient stratification.Xuan Xu et al. demonstrate how mitochondrial dysfunction serves as a unifying yet divergent element in the pathobiology of AD and glioblastoma (GBM). Single-cell analyses uncovered distinct cell type signatures, with LPAR1, regulating mitochondrial dynamics and inflammation in AD, and promoted invasion and altered bioenergetics in GBM. Further machine learning integration identified four key genes namely, EFHD1, SASH1, FAM110B, and SLC25A18 as central mitochondrial regulators. These markers show potential for improved diagnosis and therapeutic targeting although experimental validation and larger cohorts are needed to confirm their clinical relevance. Overall, single-cell deep profiling reveals key regulatory biomarkers and pathways that could improve early diagnosis and therapeutic targeting. Zhang et al. analyzed the relationship between atorvastatin and memory function by integrating data from the National Health and Nutrition Examination Survey (NHANES) and the Food and Drug Administration Adverse Event Reporting System (FAERS). Atorvastatin, a statin drug approved in 2003, is primarily proposed for treating hypercholesterolemia and preventing cardiovascular disease. NHANES findings suggested a potential protective effect of atorvastatin against memory decline while FAERS data revealed specific cognitive adverse events associated with atorvastatin use. Consequently, clinicians and patients should consider atorvastatin's potential benefits against its potential cognitive risks, accounting for individual patient variability, drug responsiveness and lifestyle factors, and implementing appropriate monitoring protocols. Also, three articles described the early screening and physical therapy-based method to improve quality of life in AD patients. Cagla Elikucuk et al. highlighted that bedside and instrumental swallowing assessments are effective for early detection of dysphagia, especially in older adults. In ICU patients, however, dysphagia is often overlooked due to cognitive and behavioral issues. Aspiration pneumonia remains a leading cause of death in this group. The authors noted that early screening and exercise-based swallowing therapy can reduce complications. They also emphasized the importance of diet modification and caregiver education as preventive strategies. Swallowing therapy may shorten hospital stays and improve outcomes while PEG tubes show no clear benefit for survival or quality of life. Malnutrition and poor functional status worsen quality of life, highlighting the need for better-designed studies on dysphagia and nutrition interventions. Wu et al. compared the effectiveness of five physiotherapy interventions in the context of AD network meta-analysis. They found that game therapy and acupuncture therapy improved mental well-being and the ability to perform daily activities, with acupuncture producing particularly notable gains in cognitive performance. In addition, music therapy and exercise therapy also contribute cognitive benefits and improvement in function outcomes, respectively. Wu et al.'s analysis demonstrated the significance of personalized physiotherapy and psychotherapeutic approaches in AD care. They also illustrated some of the therapeutic effects that were cognitive, including neuroplasticity and increased blood flow, and the reduction of amyloid protein. Overall, their findings support the use of individualized physiotherapy treatment programs to improve quality of life in AD patients. Adak et al. conducted an extensive review of AD screening models focused on early identification of at-risk individuals before clinical symptoms appear. The review encompasses a broad spectrum of in vivo and in vitro models, including pharmacologically induced and transgenic animal models, as well as neuronal cell cultures models. These models are essential for studying key AD pathophysiological mechanisms such as tau pathology, Aβ aggregation, synaptic dysfunction, and neuroinflammation. In vivo models involve behavioral...

Supplementary Figure 1 | a-b, Expression of NFAT and NFκB in reporter Jurkat cells expressing CARs or miTCRs in response to a, CD19+ Nalm6 or CD19- Molm14. b, Representative flow cytometry plot of TCR constant chain expression after dual TRAC and TRBC disruption. c, Summary data of TCR surface expression after dual TRAC and TRBC disruption in n=8 individual donors. Analysis using two-sided Student’s t-test.