bims-scepro 2026-03-08 papers

bims-scepro

Biomed News

on Stem cell proteostasis

Issue of 2026–03–08
eighteen papers selected by
William Grey, University of York

Hematopoietic stem cells activate a latent differentiation pathway to facilitate recovery after 5-fluorouracil-induced myeloablation.
Mesenchymal stromal cells modulate survival and regeneration of human hematopoietic stem cells via PGE2/cAMP signaling.
Cryopreservation and Thawing of Ex Vivo Expanded Cord Blood Hematopoietic Stem Cells.
CD49d governs immune synapse formation through actin rearrangements and synchronizes BCR signaling in CLL.
Pathological insights of human CD34+ hematopoietic stem cell-engrafted NSG mice: Implications for the safety assessment of cancer immunotherapy drugs.
Fine-Tuning of Label-Free Single-Cell Proteomics Workflows.
GCN2 in proteostasis: structural logic, signalling networks and disease.
FASN inactivation-induced progranulin (GRN) expression promotes lysosome-dependent cell death to suppress leukemogenesis.
Proteostasis meets signaling: UBE2G1 in hematopoietic stem cell aging.
Structural remodeling of the mitochondrial protein biogenesis machinery under proteostatic stress.
SPINK2 silencing suppresses leukemic proliferation and restores myeloid commitment via MECOM downregulation in acute myeloid leukaemia.
High-throughput single-cell proteomics and transcriptomics from same cells with a nanoliter-scale, spin-transfer approach.
Neighbors who talk: Mitochondria-lysosome crosstalk in homeostasis.
The molecular basis of force selectivity by PIEZO2.
Tips and tricks in cryopreservation of hematopoietic stem cells: critical points to keep under control.
Multi-view deep learning of highly multiplexed imaging data improves association of cell states with clinical outcomes.
Vascular STING activation facilitates NK cell anti-tumor immunity in small cell lung cancer.
Lipocalin 2 orchestrates resistance to ferroptosis via AXL.

Dev Cell. 2026 Mar 02. pii: S1534-5807(26)00043-2. [Epub ahead of print]

Hematopoietic stem cells activate a latent differentiation pathway to facilitate recovery after 5-fluorouracil-induced myeloablation.

Zhen Zhang, Ben Jin, Chenyu You, Ya Li, Li Lin, Jianlong Sun.

  Myeloablative chemotherapy induces hematopoietic regeneration, a process orchestrated by hematopoietic stem cells (HSCs). Although prior studies have documented enhanced HSC differentiation during this process, the temporal changes in HSC fate in response to such stress remain unclear. Here, we employed lineage tracing and mathematical modeling to investigate the cell-fate dynamics of Endothelial Protein C Receptor (EPCR)-high HSCs following 5-fluorouracil (5-FU)-induced myeloablation. Our analysis revealed a transient surge in HSC differentiation immediately after 5-FU treatment, generating primarily myeloid-biased multipotent progenitors (MPPs)-subsets that typically receive limited HSC input under steady-state conditions. Following this initial cell-fate switch, elevated HSC differentiation persisted but rapidly reverted to the homeostatic differentiation pattern observed in unperturbed hematopoiesis. Additionally, our data highlight a substantial contribution of MPPs to myeloid and lymphoid lineage regeneration following 5-FU challenge. Together, these findings delineate the sequential fate transitions adopted by HSCs during severe myeloablation and identify stage-specific differentiation patterns of HSCs in stress hematopoiesis.

Keywords:  5-fluorouracil; hematopoietic stem cell; lineage tracing; stress hematopoiesis

DOI:  https://doi.org/10.1016/j.devcel.2026.02.003
Cell Death Dis. 2026 Mar 03.

Mesenchymal stromal cells modulate survival and regeneration of human hematopoietic stem cells via PGE2/cAMP signaling.

Siva Sai Naga Anurag Muddineni, Chen Katz-Even, Adi Zipin-Roitman, Diana Rasoulouniriana, Debanjan Singha Roy, Rabeaa Aborgies, Katia Beider, Yael Raz, Neta Solomon, Gal Hershkovitz, Yael Shulman, Rony Chen, Eviatar Weizman, Claudia Waskow, Arnon Nagler, Michael Milyavsky.

Ionizing radiation and chemotherapy significantly impair hematopoietic stem and progenitor cell (HSPC) function, increasing the risk of bone marrow failure and secondary malignancies. Mesenchymal stromal cells (MSCs), critical regulators within the hematopoietic niche, maintain HSPC quiescence, self-renewal, survival, and differentiation. However, the specific pro-regenerative signaling pathways activated by MSCs in human HSPCs remain incompletely defined. Here, we show that bone marrow-derived MSCs effectively suppress irradiation-induced apoptosis and preserve the in vivo repopulation capacity of human HSPCs. Transcriptomic analysis of HSPCs revealed a pronounced upregulation of CREB target genes following MSC co-culture, consistent with increased activation of the cAMP/CREB signaling pathway. Mechanistically, MSC-secreted prostaglandin E2 (PGE2) emerged as a key mediator of cAMP induced response in HSPCs. Notably, the protective effect of Forskolin/IBMX persisted for up to 72 hours post-irradiation and significantly enhanced HSPC self-renewal. At the molecular level, we revealed reduced pro-apoptotic ASPP1 and PUMA expression, elevated p21 and stabilized anti-apoptotic MCL1 and BCL-XL proteins in human HSPCs treated with cAMP pathway agonists. Overall, our findings highlight the pivotal role of PGE2/cAMP/CREB signaling axis as a central mediator of MSC-mediated protection of human HSPCs under genotoxic stress and identify pharmacological cAMP activation as a promising strategy to protect human HSPCs against DNA damage-induced hematotoxicity.

DOI: https://doi.org/10.1038/s41419-026-08502-w
J Hematol. 2026 Feb;15(1): 23-33

Cryopreservation and Thawing of Ex Vivo Expanded Cord Blood Hematopoietic Stem Cells.

Lia Morton, Induja Arulchelvam, Chelsea McGregor, Layla Ghannouchi, Nicolas Pineault.

   Background: Umbilical cord blood (CB) is an invaluable source of hematopoietic stem and progenitor cells (HSPCs). Its use in stem cell transplantation is however constrained by the insufficient cell dose present in each unit. Recent development in ex vivo HSPC expansion technologies addresses this issue and encourages the use of the best matched CB unit. In this study, we sought to develop a cryopreservation and thawing protocol for ex vivo expanded HSPC.
Methods: CB CD34+ HSPC-enriched cells were expanded in serum-free medium supplemented with a previously optimized mix of chromatin-modifiers and early acting cytokines for 7-days. CB HSPC were then harvested and prepared for cryopreservation. Thawed CB samples were then analyzed by flow cytometry to measure cell viability and recovery of HSPC-enriched fractions, while graft potency was measured using the colony-forming unit (CFU) assay.
Results: First, we compared two widely used means of freezing; a passive isopropyl alcohol-based freezing container vs. a controlled-rate freezer (CRF). Both methods exhibited comparable recovery of viable cell numbers, including the HSC-enriched CD34+CD45RA-CD90+ fraction, and similar potency measured using the CFU assay. Next, we compared two thawing methods frequently used in clinical settings. The "thaw and dilute" method slightly improved the recovery of total nucleated cells (TNC) and HSPC fractions over the "rinse" method, though potency was comparable between both thaw methods. Next, we investigated the impact of three different commercial freezing solutions on product recovery. Dimethyl sulfoxide (DMSO)/dextran-40 and CryoProtectPure-STEM (CPP) provided superior recovery of HSPC-fractions and potency when compared to CryoScarless (CSL).
Conclusions: Taken together, this study provides insights into alternative, less harmful options for the freezing and thawing of ex vivo expanded HSPCs.

Keywords:  Cell death; Cryopreservation; Hematopoietic stem cells; Stem cell agonist cocktail; Stem cell expansion; Thawing

DOI:  https://doi.org/10.14740/jh2167
Blood. 2026 Mar 05. pii: blood.2024027753. [Epub ahead of print]

CD49d governs immune synapse formation through actin rearrangements and synchronizes BCR signaling in CLL.

Laura Polcik, Abhishek Pethe, Driti Ashok, Erika Tissino, Adrián Fernández-Rego, Federico Pozzo, Danielle-Justine Danner, Manuel Holst, Claudio Martines, Karin Hofmann, Aleksandar J Dimovski, Sandra Kissel, Andrea Härzschel, Lixia Li, Tamara Bittolo, Geoffroy Andrieux, Theresa Haslauer, Jan Philip Höpner, Nadja Zaborsky, Richard Greil, Cornelius Miething, Jesus Duque-Afonso, Natalie Köhler, Melanie Boerries, Riccardo Bomben, Justus Duyster, Robert Grosse, Gianluca Gaidano, Alberto Zamò, Antonella Zucchetto, Yolanda R Carrasco, Dimitar G Efremov, Valter Gattei, Tanja Nicole Hartmann.

B cell receptor (BCR) signaling is a key determinant of chronic lymphocytic leukemia (CLL) pathophysiology. CD49d, the alpha4 subunit of the very late antigen-4 (VLA-4) integrin, can be activated by BCR signals; however, its role in modulating BCR functionality remains unknown. We used knockout mouse models and primary human CLL stratified by CD49d expression to address this aspect. CD49d was required for bone marrow infiltration and shaped bone marrow infiltration patterns and patient outcomes in human CLL. In TCL1 transplantation models, loss of CD49d abrogated bone marrow homing and leukemic cell positioning within splenic niches. At the cellular level, CD49d-deficient murine TCL1 transgenic cells and human CD49d-low CLL cells failed to form efficient immune synapses with antigen-presenting membranes. Transcriptome analyses identified CD49d-dependent regulation of actin-associated pathways and distinct signatures of BCR responsiveness in human and mouse. Consistently, CD49d-low human CLL cells displayed aberrant actin remodeling following BCR stimulation, and a second aggressive murine CLL model reproduced the actin and engraftment defects. Kinome profiling linked impaired antigen-induced BCR responses in CD49d-deficient murine cells to altered kinase activity, and pharmacological actin perturbation phenocopied CD49d loss. In human CD49d-low CLL cells, a desynchronization of BCR-related downstream Syk and PLCɣ activation was found. Mechanistically, the CD49d-BCR interplay involved their co-localization, and CD49d converged with BCR signaling on a focal adhesion kinase-actin axis. In summary, our findings establish CD49d as a key regulator of BCR functionality in CLL, linking integrins to cytoskeletal dynamics and antigen responsiveness.

DOI: https://doi.org/10.1182/blood.2024027753
Vet Pathol. 2026 Mar 04. 3009858261423132

Pathological insights of human CD34+ hematopoietic stem cell-engrafted NSG mice: Implications for the safety assessment of cancer immunotherapy drugs.

Anna Maria Giusti, Johannes Sam, Sara Colombetti, Inês Berenguer Veiga, Josep M Monné Rodríguez.

  Humanized mice engrafted with human CD34+ hematopoietic stem cells (HSCs) are invaluable models for the preclinical evaluation of cancer immunotherapy drugs, helping to bridge the gap between in vitro studies and clinical trials. However, there are inherent challenges associated with these models, which require careful consideration, as they can obscure the distinction between treatment-related effects and model-specific background lesions. Here, we provide a comprehensive overview of common histopathological changes in such models, focusing on humanized nonobese diabetic (NOD) Cg-Prkdcscid Il2rgtm1Wjl/SzJ (huNSG) mice engrafted with human CD34+ HSCs. We characterize engraftment kinetics and histopathological changes at various timepoints post-engraftment and provide examples of huNSG mice utilized to evaluate immuno-oncology compounds and how these drugs influenced the development of huNSG-related background lesions. Flow cytometry and immunohistochemistry analyses reveal dynamic changes in human immune cell populations, with a predominance of B cells over T cells at early timepoints, shifting to T cell predominance at later stages. Notably, spontaneous granulomatous lesions, dominated by human macrophages, are observed in multiple organs, which increase in incidence and severity over time. These lesions are exacerbated by immunostimulatory treatments, highlighting the need for careful interpretation of drug-induced effects vs model-specific background pathology. Additionally, bone marrow necrosis that resembles ischemic lesions is identified in some huNSG mice, particularly following immune stimulation. Our findings underscore the critical importance of understanding the inherent pathological changes in huNSG mice to accurately assess the safety of cancer immunotherapy drugs.

Keywords:  Bone marrow necrosis; NSG mice; engraftment; granulomatous lesions; hematopoietic stem cells; humanization; immunomodulatory compounds

DOI:  https://doi.org/10.1177/03009858261423132
J Proteome Res. 2026 Mar 04.

Fine-Tuning of Label-Free Single-Cell Proteomics Workflows.

Pauline Perdu-Alloy, Charline Keller, Anjali Seth, Christine Carapito.

  Mass spectrometry-based single-cell proteomics emerges as the most promising method for studying cellular heterogeneity at the global proteome level with unprecedented depth and coverage. Its widespread application remains limited due to robustness, reproducibility, and throughput requirements, still difficult to meet as analyzing large cohorts of single cells is necessary to ensure statistical confidence. In this context, we conducted method optimizations at three levels. First, we benchmarked three distinct workflows compatible with the nanoElute2 platform using different sample collection/preparation plate supports (EVO96 oil-free, LF48 oil-based, and LF48 oil-free, a streamlined automated sample resuspension, and direct injection protocol). Then, we compared the optimized EVO96 workflow on nanoElute2 with Evosep-based separations operating at two analytical throughputs (80 and 120 samples per day). Subsequently, we evaluated digestion efficiency using a range of enzyme/protein ratios (1:1; 10:1; 20:1; 50:1) to maximize peptide recovery. Finally, the chromatographic setup was refined to determine the best compromise between throughput and robustness. Altogether, these optimizations allowed to establish a robust workflow quantifying up to 5000 proteins in 10 min gradient time per single HeLa cell at a 55 samples-per-day throughput.

Keywords:  LC–MS/MS acquisition; SCP sample preparation; dia-PASEF; digestion efficiency; label-free quantification; single-cell proteomics (SCP)

DOI:  https://doi.org/10.1021/acs.jproteome.5c01075
FEBS J. 2026 Mar 04.

GCN2 in proteostasis: structural logic, signalling networks and disease.

JiaYi Zhu, Stefan J Marciniak.

  Proteostasis is the finely tuned balance of protein synthesis, folding and degradation essential for cellular health. When this equilibrium is disrupted, misfolded proteins accumulate, triggering adaptive stress responses such as the unfolded protein response and the integrated stress response (ISR). Central to the ISR is the kinase GCN2, a sensor of amino acid deprivation and ribosomal stress. Upon activation, GCN2 phosphorylates eIF2α, dampening global translation while selectively enhancing the synthesis of the stress-responsive transcription factors ATF4 and CHOP. ATF4 orchestrates a broad transcriptional programme that supports amino acid metabolism, redox homeostasis, autophagy and proteasomal degradation, which are key processes for restoring proteostasis. Beyond its canonical role, GCN2 interfaces with other regulatory networks modulating mTORC1 to promote autophagic clearance of damaged proteins and organelles, facilitating stress granule formation, and integrating signals from oxidative and endoplasmic reticulum stress to rebalance the proteome. Dysregulated GCN2 activity has been implicated in diverse pathologies including neurodegeneration, cancer and pulmonary vascular disease, positioning it as a promising therapeutic target. In this review, we explore how GCN2 links nutrient sensing to translational control and metabolic adaptation, and how its central role in proteostasis may inform new strategies for treating diseases driven by protein misfolding and stress pathway imbalance.

Keywords:  GCN2; amino acid sensing; integrated stress response; proteostasis; translational control

DOI:  https://doi.org/10.1111/febs.70480
Cell Rep. 2026 Mar 03. pii: S2211-1247(26)00120-8. [Epub ahead of print]45(3): 117042

FASN inactivation-induced progranulin (GRN) expression promotes lysosome-dependent cell death to suppress leukemogenesis.

Meng Su, Qinglin Li, Zhiyi Lv, Xinshu Xie, Yating Lu, Yaohui He, Hanqi Liu, Hui Cao, Jie Ouyang, Qiao Zhang, Xuezhen Ma, Yexin Yang, Ailing Zou, Kang Liu, Shuqian Xu, Ji Li, Lili Chen, Xin Lu, Jie Yang, Caiming Wu, Lu Liu, Lei Zhang, Yue Sheng, Yong Huang, Yang Mei.

  Cancer cells rely on lipogenesis in addition to exogenous lipid uptake, and fatty acid synthase (FASN) is aberrantly overexpressed in myeloid leukemia, yet its role in leukemogenesis is unclear. We show that FASN is essential for leukemogenesis. Its genetic ablation impairs leukemic cell growth, survival, and clonogenicity in vitro, and reduces disease burden in vivo, without significantly affecting normal hematopoiesis. We further identify a platensimycin derivative compound MS-C19 as a potent FASN inhibitor. MS-C19 suppresses growth and clonogenicity in clinical acute myeloid leukemia (AML) samples. Mechanistically, FASN inhibition or deficiency activates lysosomal and inflammatory gene programs, inducing lysosomal membrane permeabilization and associated cell death but not lysosome biogenesis. We further identify that GRN, a lysosomal and neuroinflammatory gene, is potently transcribed by TFEB upon FASN inhibition. GRN depletion reverses the anti-leukemic effects of FASN loss. Our findings establish FASN as a therapeutic target and support its pharmacological inhibition by MS-C19 for leukemia treatment.

Keywords:  CP: cancer; fatty acid synthase; lysosome-dependent cell death; myeloid leukemia; platensimycin derivatives; progranulin

DOI:  https://doi.org/10.1016/j.celrep.2026.117042
Haematologica. 2026 Mar 05.

Proteostasis meets signaling: UBE2G1 in hematopoietic stem cell aging.

Vika Miller, Boaz Nachmias.

DOI: https://doi.org/10.3324/haematol.2026.300724
Sci Adv. 2026 Mar 06. 12(10): eaed3579

Structural remodeling of the mitochondrial protein biogenesis machinery under proteostatic stress.

Kenneth Ehses, Jorge P López-Alonso, Odetta Antico, Yannik Lang, Till Rudack, Abdussalam Azem, Miratul M K Muqit, Iban Ubarretxena-Belandia, Rubén Fernández-Busnadiego.

Cells have evolved organelle-specific responses to maintain protein homeostasis (proteostasis). During proteostatic stress, mitochondria down-regulate translation and enhance protein folding, yet the underlying mechanisms remain poorly defined. Here, we used cryo-electron tomography to observe the structural consequences of mitochondrial proteostatic stress within human cells. We detected protein aggregates within the mitochondrial matrix, accompanied by a marked remodeling of cristae architecture. Concomitantly, the number of mitochondrial ribosome complexes was significantly reduced. Mitochondrial Hsp60 (mHsp60), a key protein folding machine, underwent major conformational changes to favor complexes with its co-chaperone mHsp10. We visualized the interactions of mHsp60 with native substrate proteins and determined in vitro mHsp60 cryo-electron microscopy structures enabling nucleotide state assignment of the in situ structures. These data converge on a model of the mHsp60 functional cycle and its essential role in mitochondrial proteostasis. More broadly, our findings reveal structural mechanisms governing mitochondrial protein biosynthesis and their remodeling under proteostatic stress.

DOI: https://doi.org/10.1126/sciadv.aed3579
Cell Death Discov. 2026 Mar 03.

SPINK2 silencing suppresses leukemic proliferation and restores myeloid commitment via MECOM downregulation in acute myeloid leukaemia.

Antonio Benedetto Ventura, Tiziana Loconte, Amer Ahmed, Lucia Deligio, Antonio Negri, Gabriella D'Angelo, Daria Di Molfetta, Pierre Cauchy, Barbara Mandriani, Xiao Zhang, Crescenza Pasciolla, Antonello Rana, Angela Iacobazzi, Giacomo Loseto, Mauro Cives, Luigi Viggiano, Francesco Massimo Lasorsa, Attilio Guarini, Maria Carmela Vegliante, Sabino Ciavarella, Giancarlo Castellano, Giuseppe Fiermonte, Giacomo Volpe.

Myeloid leukaemias harboring complex karyotypes present several unrelated cytogenetic abnormalities and form a distinct subset of AML linked to a dismal prognosis. Currently, no effective options are available for the treatment of those patients, and the discovery of novel therapeutic strategies represent an urgent clinical priority. We previously developed a bioinformatic framework for the identification of novel molecular vulnerabilities for disease stratification and treatment and observed SPINK2, a serine protease inhibitor Kazal-type 2, as a novel and promising candidate target in AML, with particularly pronounced effects in complex karyotype patients. Using publicly available bulk and single cell RNA-seq datasets, we discovered a robust association between SPINK2 and cell cycle regulators, most notably S-phase genes. By performing shRNA-mediated genetic manipulation of SPINK2 expression in a complex karyotype AML cell lines, we observed a profound impairment of proliferation coupled with an induction of terminal myeloid commitment. Moreover, SPINK2-deficient FUJIOKA cells revealed a significant association between SPINK2 and MECOM expression, consistent with findings in patients harbouring complex karyotypes, yet absent in other AML subsets from the TARGET-AML cohort. Our findings suggest a novel potential correlation between SPINK2 and MECOM expression in complex karyotype leukemias and warrant further investigation into the underlying molecular mechanisms through which the SPINK2-MECOM axis enforces aberrant self-renewal and the development of novel targeted approaches aimed at modulating its expression in complex karyotypic AML.

DOI: https://doi.org/10.1038/s41420-026-02988-1
Lab Chip. 2026 Mar 06.

High-throughput single-cell proteomics and transcriptomics from same cells with a nanoliter-scale, spin-transfer approach.

Pranav Dawar, Lye Meng Markillie, Sarah M Williams, Hugh D Mitchell, Johannes W Bagnoli, Joshua Cantlon-Bruce, Anjali Seth, Carter C Bracken, Ljiljana Paša-Tolić, Ying Zhu, James M Fulcher.

Single-cell multiomic platforms provide a comprehensive snapshot of cellular states and cell types by offering critical insights into the spatiotemporal regulation of biomolecular networks at a systems level, thereby defining the basis of multicellularity. Here, we introduce nanoSPINS, an advanced platform that enables high-throughput profiling and integrative analysis of the transcriptome and proteome from the same single cells using RNA sequencing and isobaric labeling LC-MS-based proteomics, respectively. NanoSPINS can efficiently transfer mRNA-containing droplets across two microarrays via a centrifugation-based approach, while proteins are retained on the initial platform. Benchmarking of nanoSPINS on two cell lines demonstrates its ability to generate global proteomic and transcriptomic profiles that align well with previously established methodologies/platforms. The incorporation of isobaric TMTpro labeling into this single-cell multiomics platform significantly enhances the throughput of single-cell proteomic analyses. Through the high-throughput quantification of the proteome and transcriptome, nanoSPINS not only facilitates the identification of molecular features at both mRNA and protein level but also provides larger sample sizes for improved statistical power in clustering and differential abundance. Given the broad applicability of single-cell multiomics in biological research and clinical settings, we believe nanoSPINS represents a powerful platform for the characterization of heterogeneous cell populations.

DOI: https://doi.org/10.1039/d5lc01008j
Curr Opin Cell Biol. 2026 Mar 05. pii: S0955-0674(26)00015-3. [Epub ahead of print]100 102627

Neighbors who talk: Mitochondria-lysosome crosstalk in homeostasis.

Akriti Prashar, Rosa Puertollano.

Mitochondria are highly dynamic and multifaceted organelles that perform essential cellular functions such as producing energy, regulating metabolism, and orchestrating immune responses. Lysosomes are crucial signaling hubs that are important for nutrient sensing, signal transduction, and regulation of cellular degradation and recycling processes including the removal of damaged mitochondrial components or entire mitochondria. Together, these two organelles perform critical cellular functions. Emerging evidence links defects in both organelles to multiple diseases, underscoring how their functions are intricately linked. To coordinate their activities, mitochondria and lysosomes engage in bidirectional crosstalk, enabling reciprocal regulation of their respective functions. These 'organelle conversations' can occur through direct interactions at membrane contact sites where both organelles physically interact via stabilization by molecular tethers, or at a distance through signaling pathways. Here we discuss recent progress in our understanding of the mechanisms underlying mitochondria-lysosome crosstalk and how this communication is altered in pathological conditions.

DOI: https://doi.org/10.1016/j.ceb.2026.102627
Nature. 2026 Mar 04.

The molecular basis of force selectivity by PIEZO2.

Eric M Mulhall, Oleg Yarishkin, Rose Z Hill, Anna K Koster, Ardem Patapoutian.

PIEZOs are mechanically gated ion channels that transduce force into electrochemical signals1. PIEZO1 responds to diverse stimuli including membrane stretch2 and shear stress3, whereas PIEZO2 is generally tuned to detect cellular indentation4,5. The functional specialization of PIEZO2 is proposed to underlie its distinct physiological roles, including mediating the sense of touch6,7. How PIEZO2 achieves this selectivity despite its close structural similarity to PIEZO1 is unclear. Here we combine single-molecule MINFLUX fluorescence nanoscopy with electrophysiology to link the conformational states of PIEZO2 to channel gating in intact cells. We find that PIEZO2 is intrinsically more rigid than PIEZO1, and that disparate mechanical stimuli paradoxically evoke opposite conformational and gating responses in each channel. These unique gating properties arise in part from a connection to the actin cytoskeleton, and we identify filamin-B (FLNB) as a molecular tether that is required for this interaction. This complex alters how force is transmitted to PIEZO2 and confers heightened sensitivity to and selectivity for cellular indentation. PIEZO2 and FLNB are co-expressed in somatosensory neurons and colocalize within tens of nanometres at the end organs of cutaneous mechanosensory afferents. These findings help to explain why PIEZO2 is a specialized mechanosensor and provide a molecular blueprint for understanding how cells decode diverse mechanical stimuli across tissues and organ systems.

DOI: https://doi.org/10.1038/s41586-026-10182-7
Cytotherapy. 2026 Jan 03. pii: S1465-3249(26)00001-0. [Epub ahead of print]28(5): 102045

Tips and tricks in cryopreservation of hematopoietic stem cells: critical points to keep under control.

Ivana Ferrero, Deborah Rustichelli, Sara Castiglia, Alessandra Mandese, Aloe Adamini, Marco De Gobbi, Alessandro Cignetti, Beatrice Sani, Francesco Saglio, Franca Fagioli.

   BACKGROUND AIMS: Several studies demonstrated that under standard conditions, CD34+ hematopoietic stem cells remained viable after cryopreservation for up to 19 years. However, to ensure high-quality cellular products, it is essential to define control points throughout the HSC manipulation process and validate any critical procedure. The main aim of this study is the description of the validation plan used to analyze the cryopreservation procedure and its correlation with the clinical outcome of the procedure.
METHODS: A validation plan was defined based on an FMEA risk assessment used to identify critical control points of the whole process. A careful risk analysis allows keeping the critical aspects of the process under control to guarantee patient safety.
RESULTS: Data about neutrophil and platelet engraftment confirmed the impact of the validation process on the success of the transplantation procedure. We also analyzed CD34+ viability in relation to total nucleated cells (TNC) concentration (cells/mL) in cryopreserved cellular products collected over the past three years (2023-2025).
CONCLUSIONS: This work provides a practical guide for operators, offering clear indications on how to manage and control each critical step of the HSC cryopreservation process to ensure product quality and patient safety.

Keywords:  cryopreservation; hematopoietic stem cell transplantation; risk analysis; validation

DOI:  https://doi.org/10.1016/j.jcyt.2026.102045
Bioinform Adv. 2026 ;6(1): vbag010

Multi-view deep learning of highly multiplexed imaging data improves association of cell states with clinical outcomes.

Shanza Ayub, Jennifer L Gorman, Edward L Y Chen, Hartland W Jackson, Alina Selega, Kieran R Campbell.

Motivation: Analysis workflows for highly multiplexed imaging technologies typically summarize each cell in terms of its post-segmentation mean expression, but additional cellular information can be quantified including cell morphology, sub-cellular expression patterns, and spatial cellular context, ultimately giving a multi-modal view of each cell. While deep learning models such as variational autoencoders are well-established for other multi-modal single-cell assays, their ability to integrate these multiple views of a cell from highly multiplexed imaging data remains largely unknown.
Results: Here, we explore the abilities of multi-modal variational autoencoders to learn unified latent cellular representations from multiple views of each single-cell quantified from highly multiplexed imaging, including mean expression, morphology, sub-cellular protein co-localization, and spatial cellular context, while conditioning on technical and batch specific effects. We show that the integrated multi-modal latent space is often more associated with patient-specific clinical outcomes compared to a set of existing baselines. In addition, we perform ablation analyses to understand which input views contribute to model performance, and explore the ability of these models to learn cellular representations that align with cellular phenotypes and enable integration across divergent datasets.
Availability and implementation: hmiVAE is implemented as a python package and is available at https://github.com/camlab-bioml/hmiVAE.

DOI: https://doi.org/10.1093/bioadv/vbag010
Cancer Cell. 2026 Mar 05. pii: S1535-6108(26)00106-6. [Epub ahead of print]

Vascular STING activation facilitates NK cell anti-tumor immunity in small cell lung cancer.

Marco Campisi, Tatsuya Osaki, Ian Dryg, Carla Stornante, Jacqueline Wolff, Jason Weirather, Nicholas Weaver, Mubin Tarannum, Ian Gillanders, Alan Bers, Eden Bobilev, Maia S Lineberry, Pieter Schol, Minyue Chen, Keiichi Ota, Sung R Park, Caroline G Fahey, Tran C Thai, Yixiang Li, Zhaorong Li, Ze-Hua Li, Sarah Shelton, Satomi Hirose, Clare Padrick, Elena Ivanova, Minh Ha, Shaobo Yang, Harrison Olszewski, Sophie Kivlehan, Patrick Lizotte, Kelsey Hagen, Iliana Gjeci, William Haller, Lauren M Zasadil, Talal El Zarif, Jacob E Berchuck, Erik H Knelson, Elliott J Brea, Igor Odintsov, Tianju Liu, Valeria Chiono, Bishma Tuladhar, Kenneth Ngo, Emily Rosentrater, Jeffrey Raizer, Neil Lineberry, Mriganka Sur, Kathleen Pfaff, Matthew Freedman, F Stephen Hodi, Michael Y Tolstorukov, Matthew G Oser, Michal Sheffer, Constantine S Mitsiades, Michael J Barnes, Vicky A Appleman, Prafulla C Gokhale, Roger D Kamm, Cloud P Paweletz, Rizwan Romee, Scott Rodig, David A Barbie, Navin R Mahadevan.

  Small cell lung cancer (SCLC) typically displays a "cold" tumor microenvironment with a paucity of immune infiltrate. Neuroendocrine SCLC cells also profoundly repress MHC-I expression, rendering them vulnerable to NK cell-mediated cytotoxicity. Here, we confirm that neuroendocrine SCLC cells are sensitive to NK cell-mediated attack, yet the quantitative spatial profiling of the SCLC immune microenvironment in patient samples reveals that effector immune cells, including NK cells, are excluded from MHC-Ilow/neg SCLC regions. To study this biology, we develop dynamic single-cell RNA sequencing of microphysiological immune tumor environments (DynaMITE-seq) and integrate findings with spatial transcriptomics in patient tissue, unveiling the microvasculature as a major checkpoint restricting NK cell extravasation/recruitment. We demonstrate that the activation of vascular Stimulator of Interferon Genes (STING) signaling restores NK cell infiltration and killing of neuroendocrine SCLC, suggesting a strategy to overcome this key SCLC immunologic barrier and prime therapeutic response to DLL3-targeted CAR-NK cell therapy.

Keywords:  3D microphysiological systems; STING agonism; natural killer cells; small cell lung cancer; spatial transcriptomics and proteomics; tumor-immune microenvironment

DOI:  https://doi.org/10.1016/j.ccell.2026.02.008
Cell Rep. 2026 Feb 27. pii: S2211-1247(26)00043-4. [Epub ahead of print]45(3): 116965

Lipocalin 2 orchestrates resistance to ferroptosis via AXL.

Sabrina Z Wang, J Payton Timken, Ellen S Hong, Sehaj Kaur, Eli Newby, Kristen E Kay, Erin E Mulkearns-Hubert, Daniel J Silver, Juyeun Lee, Joshua B Rubin, James R Connor, Loic P Deleyrolle, Deanna Tiek, Andrew Dhawan, Justin D Lathia.

  Glioblastoma (GBM) remains a lethal tumor, largely due to robust mechanisms that prevent effective induction of cell death. Ferroptosis, a form of iron-dependent cell death, is a promising vulnerability in GBM. Here, we demonstrate that lipocalin-2 (LCN2) suppresses ferroptosis in GBM cells via the receptor tyrosine kinase AXL. LCN2 was elevated in GBM cells compared to lower-grade tumor and non-transformed cells, and Lcn2 knockdown impaired GBM cell fitness and growth in vitro and in vivo. Mechanistically, Lcn2 knockdown triggered ferroptosis, which was specifically rescued with ferroptosis inhibitors but not apoptosis or necroptosis inhibitors. Lcn2 knockdown reduced AXL phosphorylation, which was elevated in GBM patient tumors relative to non-tumor tissue. Notably, the combination of Lcn2 knockdown and pharmacological AXL inhibition extended survival compared to Lcn2 knockdown alone. Taken together, these data reveal a link between LCN2-mediated suppression of ferroptosis with AXL phosphorylation and support this axis as a potential therapeutic target for GBM.

Keywords:  AXL; CP: cancer; cell death; glioblastoma; lipocalin-2

DOI:  https://doi.org/10.1016/j.celrep.2026.116965