bims-cagime Biomed News
on Cancer, aging and metabolism
Issue of 2025–06–08
35 papers selected by
Kıvanç Görgülü, Technical University of Munich



  1. bioRxiv. 2025 May 20. pii: 2025.05.15.654370. [Epub ahead of print]
      Cancer cachexia is an involuntary weight loss condition characterized by systemic metabolic disorder. A comprehensive flux characterization of this condition however is lacking. Here, we systematically isotope traced eight major circulating nutrients in mice bearing cachectic C26 tumors (cxC26) and food intake-matched mice bearing non-cachectic C26 tumors (ncxC26). We found no difference in whole-body lipolysis and proteolysis, ketogenesis, or fatty acid and ketone oxidation by tissues between the two groups. In contrast, compared to ncxC26 mice ad libitum, glucose turnover flux decreased in food intake-controlled ncxC26 mice but not in cxC26 mice. Similarly, sustained glucose turnover flux was observed in two autochthonous cancer cachexia models despite reduced food intake. We identified glutamine and alanine as responsible for sustained glucose production and tissues with altered use of glucose and lactate in cxC26 mice. We provide a comprehensive view of metabolic alterations in cancer cachexia revealing those distinct from decreased nutrient intake.
    Highlights: Quantitative fluxomics of cancer cachexia under matched food intake and body weightIntact lipolysis, proteolysis, ketogenesis, and lipid oxidation in cachectic miceSustained glucose consumption in cachectic mice despite reduced food intakeIncreased glucose production from glutamine and alanine in cachectic mice.
    DOI:  https://doi.org/10.1101/2025.05.15.654370
  2. bioRxiv. 2025 May 24. pii: 2025.05.20.655200. [Epub ahead of print]
      Pancreatic ductal adenocarcinoma (PDAC) is an aggressive cancer with poor outcomes. Obesity is a risk factor for several cancers including PDAC due to metabolic dysregulation and inflammation. The ketogenic diet (KD) can alter metabolism and has been evaluated for its effects on tumor progression in non-obese but not obese PDAC using genetically engineered mouse models (GEMMs). We hypothesized that ketone bodies and a KD alter cell and tumor metabolism. We show that ketone treatments altered pyrimidine metabolism in PDAC cells. Moreover, in an obese PDAC GEMM, KD prevented tumor progression independent of weight loss but promoted PDAC in a non-obese PDAC GEMM. The KD-specific delay of obesity-associated PDAC was associated with pancreatic metabolic shifts in pyrimidine, cysteine and methionine, and arginine and proline pathways. These findings suggest potential benefits of a KD in preventing obesity-associated PDAC, but highlights some risks in non-obese settings.
    DOI:  https://doi.org/10.1101/2025.05.20.655200
  3. Gut. 2025 Jun 04. pii: gutjnl-2025-335439. [Epub ahead of print]
      
    Keywords:  LIPID METABOLISM; PANCREATIC CANCER
    DOI:  https://doi.org/10.1136/gutjnl-2025-335439
  4. J Phys Chem B. 2025 Jun 05.
      Giant membrane vesicles (GUVs) and giant plasma membrane vesicles (GPMVs) are valuable models for studying the properties of cellular membranes. We analyzed experimental data on vesicle shapes in three-dimensional space to estimate their reduced volumes, focusing on osmotic deflation and membrane asymmetry. Shape changes in GPMVs illustrate how osmolarity influences the membrane structure in the absence of the cytoskeleton or other cellular organelles. By examining the experimentally observed shapes and their corresponding reduced volumes, we compared GPMV shapes to theoretical predictions for simple phospholipid vesicles, utilizing the area-difference elasticity and spontaneous curvature models. We mapped DOPC GUVs using the area-difference elasticity model and applied the spontaneous curvature model to map DOPC: cholesterol GUVs and GPMVs. The reported experiments showcase advanced methods that provide valuable biophysical insights, demonstrating that the GPMV shape observed in the experiments and their reduced volume can be mapped onto the same shape diagram as red blood cells (RBCs) and vesicles composed of phospholipids. This finding offers new perspectives in the field.
    DOI:  https://doi.org/10.1021/acs.jpcb.4c07431
  5. Nat Metab. 2025 Jun 03.
      Caloric restriction and methionine restriction-driven enhanced lifespan and healthspan induces 'browning' of white adipose tissue, a metabolic response that increases heat production to defend core body temperature. However, how specific dietary amino acids control adipose thermogenesis is unknown. Here, we identified that weight loss induced by caloric restriction in humans reduces thiol-containing sulfur amino acid cysteine in white adipose tissue. Systemic cysteine depletion in mice causes lethal weight loss with increased fat utilization and browning of adipocytes that is rescued upon restoration of cysteine in diet. Mechanistically, cysteine-restriction-induced adipose browning and weight loss requires sympathetic nervous system-derived noradrenaline signalling via β3-adrenergic-receptors that is independent of FGF21 and UCP1. In obese mice, cysteine deprivation induced rapid adipose browning, increased energy expenditure leading to 30% weight loss and reversed metabolic inflammation. These findings establish that cysteine is essential for organismal metabolism as removal of cysteine in the host triggers adipose browning and rapid weight loss.
    DOI:  https://doi.org/10.1038/s42255-025-01297-8
  6. Cell Rep Med. 2025 May 28. pii: S2666-3791(25)00220-4. [Epub ahead of print] 102147
      Heart failure can be worse when associated with obesity, elevated serum pancreatic enzymes, elevated non-esterified fatty acids (NEFAs), or acute pancreatitis (AP). To understand this, here we study doxorubicin-induced heart failure, experimental AP, or pancreatic lipase-induced visceral fat necrosis in lean, genetically obese (ob/ob), or dual ob/ob pancreatic triglyceride lipase (PNLIP)-knockout mice. NEFA generation and resulting cardiac injury are measured. We note that ob/ob mice develop fat necrosis containing PNLIP and phospholipase A2. This generates excess NEFAs that worsen cardiac injury, cause hypotension, and reduce survival. All these are prevented by PNLIP deletion or pharmacologic inhibition. Live imaging shows that phospholipase A2 damages adipocyte membranes, resulting in PNLIP entry and leakage of adipocyte lipases. PNLIP hydrolyzes adipose triglyceride, generates NEFAs, and causes lipid droplet loss and adipocyte necrosis. Therefore, pancreatic injury can worsen antecedent heart failure by leaked PNLIP, causing excessive visceral adipose lipolysis. Inhibition of such lipolysis may improve heart failure outcomes.
    Keywords:  adipose; cell death; fat; fatty acids; heart failure; lipase; lipid; mortality; necrosis; triglyceride
    DOI:  https://doi.org/10.1016/j.xcrm.2025.102147
  7. J Clin Invest. 2025 Jun 02. pii: e193205. [Epub ahead of print]135(11):
      Cell plasticity is a hallmark of cancer, enabling tumor cells to acquire multiple phenotypes responsible for tumor progression, metastasis, and therapy resistance. In this issue of the JCI, Kawai and colleagues leveraged genetically engineered mouse models (GEMM) of pancreatic ductal adenocarcinoma (PDAC) to demonstrate that loss of Pbrm1, a member of the SWI/SNF complex, drives dedifferentiation and aggressive tumor features. Pbrm1 loss activated a program of epithelial-to-mesenchymal transition (EMT) and allowed the emergence of poorly differentiated histologies that are commonly associated with high recurrence rate and dismal prognosis. These findings reveal the role of the SWI/SNF complex during PDAC evolution in maintaining cell identity and restraining the progression of this lethal disease.
    DOI:  https://doi.org/10.1172/JCI193205
  8. bioRxiv. 2025 May 13. pii: 2025.05.10.653256. [Epub ahead of print]
      Cancer cells alter their mechanical properties in response to the rigidity of their environment. Here, we explored the implications of this environmental mechanosensing for anti-tumor immunosurveillance using single cell biophysical profiling and metastasis models. Cancer cells stiffened in more rigid environments, a biophysical change that sensitized them to cytotoxic lymphocytes. In immunodeficient mice, this behavior manifested in the outgrowth of stiffer metastatic cells in the rigid bone than in the soft lung, while in immunocompetent hosts, it led to preferential elimination of stiffer cancer cells and suppression of bone metastasis. Environmentally-induced cell stiffening and immune sensitization both required Osteopontin, a secreted glycoprotein that is upregulated during bone colonization. Analysis of patient metastases spanning mechanically distinct tissues revealed associations between environmental rigidity, immune infiltration, and cancer cell stiffness consistent with mechanically driven immunosurveillance. These results demonstrate how environmental mechanosensing modulates anti-tumor immunity and suggest a mechanoimmunological basis for metastatic site selection.
    DOI:  https://doi.org/10.1101/2025.05.10.653256
  9. Nat Genet. 2025 Jun 02.
      To understand genetic evolution in cancer during metastasis, we analyzed genomic profiles of 3,732 cancer patients in whom several tumor sites were longitudinally biopsied. During distant metastasis, tumors were observed to accumulate copy number alterations (CNAs) to a much greater degree than mutations. In particular, the development of whole genome duplication was a common event during metastasis, emerging de novo in 28% of patients. Loss of 9p (including CDKN2A) developed during metastasis in 11% of patients. To a lesser degree, mutations and allelic loss in human leukocyte antigen class I and other genes associated with antigen presentation also emerged. Increasing CNA, but not increasing mutational load, was associated with immune evasion in patients treated with immunotherapy. Taken together, these data suggest that CNA, rather than mutational accumulation, is enriched during cancer metastasis, perhaps due to a more favorable balance of enhanced cellular fitness versus immunogenicity.
    DOI:  https://doi.org/10.1038/s41588-025-02204-3
  10. J Clin Invest. 2025 Jun 02. pii: e177533. [Epub ahead of print]135(11):
      Mutations in Polybromo 1 (PBRM1), a subunit of the switch/sucrose nonfermentable (SWI/SNF) chromatin remodeling complex, are frequently observed in several cancers, including pancreatic ductal adenocarcinoma (PDAC). In this study, we demonstrated that pancreas-specific loss of Pbrm1 in mice harboring Kras mutations and Trp53 deletions accelerated the development of poorly differentiated PDAC, epithelial-mesenchymal transition (EMT), and metastasis, resulting in worsened prognosis. Pbrm1 loss in preexisting PDAC shifted the tumor grade from a well- to a poorly differentiated state and elevated vimentin expression. Pbrm1-null PDAC exhibited downregulation of apical junction genes and upregulation of EMT pathway genes, including the vimentin and squamous molecular subtype signature genes. Mechanistically, PBRM1 bound to the vimentin gene promoter and directly downregulated its expression. Furthermore, suppression of vimentin in Pbrm1-null PDAC cells reversed the dedifferentiation phenotype and reduced EMT and metastasis. Consistently, reduced PBRM1 expression correlated with high vimentin expression, poorly differentiated histology, a high recurrence rate, and reduced overall survival in human PDACs. Additionally, PDAC with PBRM1 deletion was associated with the aggressive squamous molecular subtype. Our data established PBRM1 as a tumor suppressor that controls tumor grade and metastasis of PDAC by regulating vimentin expression.
    Keywords:  Cancer; Epigenetics; Gastroenterology; Mouse models; Oncology
    DOI:  https://doi.org/10.1172/JCI177533
  11. Proc Natl Acad Sci U S A. 2025 Jun 10. 122(23): e2425347122
      Metabolism fuels cell growth and functions. While it is well established that cellular growth rate scales with cell size, how cells alter their metabolism as they change size remains largely unexplored. Here, we conducted a systematic analysis of cell size-dependent metabolism across the NCI60 cancer cell line panel comprising a diverse range of cell sizes. We demonstrate that cellular metabolism and growth rate display 2/3 allometric scaling due to differential scaling of overall nutrient uptake and waste metabolite release with respect to cell size, with waste elimination decreasing less rapidly than nutrient uptake rate as cells grow larger. This results in cell size-dependent growth rate and predicts a maximum cell size where net nutrient uptake equals zero and cell enlargement ceases despite active metabolism. We experimentally confirm this prediction and identify that electron acceptor demand constrains cell enlargement as evidenced by depletion of intracellular aspartate and scaling of aspartate uptake, which is more than proportional to cell volume. Overall, these findings may have implications for understanding cell size homeostasis, developmental biology, and the design principles of living organisms.
    Keywords:  allometry; cell size; growth rate; metabolism; scaling
    DOI:  https://doi.org/10.1073/pnas.2425347122
  12. Dev Cell. 2025 May 27. pii: S1534-5807(25)00318-1. [Epub ahead of print]
      Lysosome damage activates multiple pathways to prevent lysosome-dependent cell death, including a repair mechanism involving endoplasmic reticulum (ER)-lysosome membrane contact sites, phosphatidylinositol 4-kinase-2a (PI4K2A), phosphatidylinositol-4 phosphate (PI4P), and oxysterol-binding protein-like proteins (OSBPLs) lipid transfer proteins. PI4K2A localizes to the trans-Golgi network and endosomes, yet how it is delivered to damaged lysosomes remains unknown. During acute sterile damage and damage caused by intracellular bacteria, we show that ATG9A-containing vesicles perform a critical role in delivering PI4K2A to damaged lysosomes. ADP ribosylation factor interacting protein 2 (ARFIP2), a component of ATG9A vesicles, binds and sequesters PI4P on lysosomes, balancing OSBPL-dependent lipid transfer and promoting the retrieval of ATG9A vesicles through the recruitment of the adaptor protein complex-3 (AP-3). Our results identify a role for mobilized ATG9A vesicles and ARFIP2 in lysosome homeostasis after damage and bacterial infection.
    Keywords:  AP-3; ARFIP2; ATG9A; PI4K2A; PI4P; autophagy; lysosomal damage; lysosome; membrane trafficking
    DOI:  https://doi.org/10.1016/j.devcel.2025.05.007
  13. Biochemistry. 2025 Jun 04.
      Selenoprotein K (selenok) is a small, disordered membrane protein associated with the endoplasmic reticulum (ER) that is involved in protein palmitoylation and protein quality control. Through these processes, it influences calcium homeostasis, cellular migration, and phagocytosis. Thus, it is implicated in cancer, neurodegenerative diseases, and autophagy. So far, selenok has been considered a single-pass membrane protein whose N-terminus is in the ER lumen while its C-terminus, which contains the reactive selenocysteine, is in the cytoplasm. Here, we show that selenok is, in fact, a peripheral membrane protein that is anchored to the cytoplasmic side of the ER membrane. We demonstrate, using immunofluorescence microscopy and the substituted cysteine accessibility method in combination with selective membrane permeabilization, that both selenok's N- and C-terminus are in the cytoplasm. Using the same techniques, we demonstrate that, in contrast, selenoprotein S (selenos), a functionally related member of the selenoprotein family, is a transmembrane protein with a cytoplasmic C-terminus and an N-terminus exposed to the ER lumen. The findings that selenok is a peripheral membrane protein and that its N- and C-terminal segments, along with the hydrophilic side of its amphipathic α-helix, are exposed to the cytoplasm, imply that they can interact with cytoplasmic extramembranous regions of ER-residing membrane proteins and soluble protein partners. Selenok is predicted to possess multiple SLiMs (short linear motifs) involved in protein interactions, and its peripheral topology suggests that all these motifs, including those located within the amphipathic α-helix, are exposed and accessible to cytoplasmic-accessible partners.
    DOI:  https://doi.org/10.1021/acs.biochem.5c00062
  14. Cell Rep. 2025 Jun 04. pii: S2211-1247(25)00560-1. [Epub ahead of print]44(6): 115789
      Caveolae are specialized plasma membrane domains with a unique lipid composition. Lipid peroxidation has recently been implicated in triggering caveola disassembly, releasing cavin proteins to regulate oxidative-stress-associated cellular processes, particularly ferroptosis. Here, we investigated how specific lipids influence caveola formation and their response to oxidative stress. A targeted screening of pro-ferroptotic enzymes identified ACSL4, a key enzyme in synthesizing polyunsaturated fatty acid (PUFA)-linked phospholipids, and ether phospholipid biosynthesis enzymes as critical regulators of caveola formation. Membrane-incorporated omega-6 PUFAs promoted caveola formation, while their displacement by omega-3 PUFAs or monounsaturated fatty acids disrupted this process. Importantly, oxidation of omega-6 PUFA chains in phosphatidylethanolamine (PE) triggered caveola disassembly during lipid peroxidation, potentially by affecting cavin-membrane interactions. These findings unveil a new model for caveola formation and signaling, linking caveola dynamics to ferroptosis with pro-ferroptotic lipids as essential caveolar components and key control points for caveola disassembly under oxidative stress.
    Keywords:  ACSL4; CP: Cell biology; MUFA; PUFA; cCaveolae; fFerroptosis; lLipids; pPlasmalogens
    DOI:  https://doi.org/10.1016/j.celrep.2025.115789
  15. Nat Rev Mol Cell Biol. 2025 Jun 05.
      Migrating cells have key functions in shaping tissues during development, repairing tissues after development and supporting cancer invasion and metastasis. In all these contexts, cells often maintain contact with their neighbours and move as a group, in a process termed collective migration. In this Review, we describe the elegant mechanisms used by collectively migrating cells in vivo to coordinate their movements and obtain directional information. We start by highlighting the diverse physiological roles that migrating collectives have within the body and then focus on dominant paradigms for the organization of migrating collectives including the roles of leader and follower cells, local cell-cell adhesion and signalling, and external guidance cues. By comparing collective migrations occurring during development and cancer, we bring into focus shared principles for collective cell movement and distinct strategies used by cancer cells for their own dispersal. Throughout, we pay particular attention to how migrating collectives display emergent properties not exhibited by individually migrating cells and how these properties provide the robustness needed for efficient cell movement.
    DOI:  https://doi.org/10.1038/s41580-025-00858-9
  16. FEBS Lett. 2025 May 31.
      Cryo-electron microscopy has yielded high-resolution structural data of the multidrug efflux transporter P-glycoprotein (ABCB1), but its direct and indirect interactions within the native membrane environment have remained largely unexplored. Here, we compared the fluidity gradients of plasma membranes of the drug-sensitive CHO cell line AuxB1 and its P-glycoprotein overexpressing derivative B30 by fluorescence anisotropy of embedded n-(9-anthroyloxy) fatty acid probes (n = 2, 7, 9, 12, 16) in the temperature range of 10-50 °C. The shape of the temperature profiles of probe mobility was comparable in AuxB1 and B30 membranes, but did not match. Overexpression of P-glycoprotein smoothened the transversal gradient of the out-of-plane mode of rotation of the probes, which may facilitate the partitioning of hydrophobic drugs into the membrane and thereby increase the speed of P-glycoprotein to pump the drug out of the cell.
    Keywords:  ABC transporter; ABCB1; P‐glycoprotein; membrane fluidity; multidrug resistance; n‐(9‐anthroyloxy) fatty acid
    DOI:  https://doi.org/10.1002/1873-3468.70083
  17. Acta Physiol (Oxf). 2025 Jul;241(7): e70067
       AIM: Cancer cachexia is a severe metabolic disorder leading to skeletal muscle atrophy. Muscle wasting is a major clinical problem in cachectic patients, as it limits the efficacy of chemotherapeutic treatments and worsens quality of life. Nutritional support based on branched-chain amino acids (BCAA) has been shown to be a promising approach to counteract cachexia-induced muscle atrophy, but its efficacy is still debated. Furthermore, the putative role of di-alanine (Di-Ala) supplementation has yet to be evaluated. The present study therefore sought to assess whether BCAA supplementation, alone or in combination with a Di-Ala peptide, could attenuate muscle wasting in a preclinical model of cancer cachexia.
    METHODS: To this end, C26 tumor-bearing mice were administered BCAA supplementation, with or without Di-Ala. Body and muscle weights, as well as molecular, biochemical, and morphological analysis, were carried out to characterize prospective changes of markers involved in cachexia and muscle atrophy.
    RESULTS: The main findings revealed that BCAA supplementation effectively prevented body weight loss and muscle atrophy. Of note, Di-Ala significantly amplified the effects of BCAA. These phenomena were found to be mediated by the suppression of pathways involved in protein catabolism.
    CONCLUSIONS: Collectively, these results highlight that innovative formulations containing Di-Ala, capable of increasing BCAA bioavailability, may be efficacious in counteracting muscle atrophy, especially during mild-to-moderate cancer cachexia.
    Keywords:  BCAA; dietary supplements; muscle wasting; nutraceuticals; protein catabolism; skeletal muscle
    DOI:  https://doi.org/10.1111/apha.70067
  18. Int J Cancer. 2025 Jun 02.
      Pancreatic cancer patients with cachexia experience functional changes in visceral organs. To further understand these functional changes, here, for the first time, we characterized metabolic changes in the spleen, liver, pancreas, lungs, heart, and kidneys induced by human pancreatic cancer xenografts. These studies identify the commonality and consequences of cachexia-induced visceral organ metabolic dysregulation. The heart, kidneys, liver, lungs, pancreas, and spleen from euthanized non-tumor-bearing control mice and from cachexia-inducing Pa04C and non-cachexia-inducing Panc1 tumor-bearing mice (n = 8-10 per group) were metabolically characterized with 1H magnetic resonance spectroscopy. All visceral organs, with the exception of lungs, exhibited significant weight reduction in cachectic Pa04C mice relative to normal and non-cachectic Panc1 mice. A significant reduction (p ≤ .0166) of organ metabolites ranging from the amino acids leucine, isoleucine, valine, alanine, lysine, arginine, asparagine, glutamate, glutamine, aspartate, glycine, tyrosine, and phenylalanine, along with glucose, lactate, creatine, choline, and fumarate, depending upon the visceral organ, was observed in cachectic Pa04C mice compared to normal mice. The highest number of metabolites was reduced in the spleen, followed by the kidneys, lungs, and liver. The metabolic changes identified can lead to negative consequences in organ function by impacting pathways involved in tissue regeneration and resolving inflammation at the cellular level in cachectic mice. These results highlight the visceral organ metabolic reprogramming that can occur with cancer-induced cachexia, an understanding of which can identify noninvasive biomarkers and metabolic interventions to reduce morbidity and mortality from pancreatic cancer.
    Keywords:  1H MR spectroscopy; cachexia; human pancreatic cancer xenografts; metabolites
    DOI:  https://doi.org/10.1002/ijc.35487
  19. bioRxiv. 2025 May 16. pii: 2025.05.13.653868. [Epub ahead of print]
      The diversity of cellular and tissue structures can arise from a few basic cell shapes, which undergo various transformations based on biophysical constraints on cytoskeletal organization. While cellular geometry has been linked with selected biological processes such as polarity, signaling or morphogenesis, the orchestration of the whole proteome in association to cell shape is still poorly understood. In this study, using more than 1 million images of single cells stained for 11,998 proteins across 10 cell lines in the Human Protein Atlas database, we performed an integrated analysis of organelle, pathway and single protein levels in association to a 2D cellular shapespace. We found that cell and nuclear shapes across cell lines exist in a shared continuum. We also found that the subcellular organelle topology varies across cell lines, but remains robust within each cell line's shapespace. At the single protein level, we found that cells of different shapes in the same cell cycle phase might be preparing for different fates, and that many non-cell cycle proteins expressed shape-based abundance variation. Using the same coordinate framework defined by shape, we could analyze the distribution shift of protein spatial localization under drug perturbation.
    DOI:  https://doi.org/10.1101/2025.05.13.653868
  20. Trends Mol Med. 2025 Jun 02. pii: S1471-4914(25)00115-7. [Epub ahead of print]
      Neuropsychiatric complications of cancer cachexia include apathy. Zhu et al. identified a brain circuit sensitive to tumor-driven interleukin-6 (IL-6) signaling that directly reduces motivation by suppressing dopaminergic activity. Targeted circuit-based and pharmacologic interventions, including systemic anti-IL-6 antibodies, reversed motivational deficits and point to new therapeutic avenues for apathy.
    Keywords:  IL-6; apathy; brain–body; cancer cachexia; metabolism; neuroimmune circuit
    DOI:  https://doi.org/10.1016/j.molmed.2025.05.004
  21. Nat Commun. 2025 May 31. 16(1): 5059
      Metabolic reactions play important roles in organisms such as providing energy, transmitting signals, and synthesizing biomacromolecules. Charting unknown metabolic reactions in cells is hindered by limited technologies, restricting the holistic understanding of cellular metabolism. Using mass spectrometry-resolved stable-isotope tracing metabolomics, we develop an isotopologue similarity networking strategy, namely IsoNet, to effectively deduce previously unknown metabolic reactions. The strategy uncovers ~300 previously unknown metabolic reactions in living cells and mice. Specifically, we elaborately chart the metabolic reaction network related to glutathione, unveiling three previously unreported reactions nestled within glutathione metabolism. Among these, a transsulfuration reaction, synthesizing γ-glutamyl-seryl-glycine directly from glutathione, underscores the role of glutathione as a sulfur donor. Functional metabolomics studies systematically characterize biochemical effects of previously unknown reactions in glutathione metabolism, showcasing their diverse functions in regulating cellular metabolism. Overall, these newly uncovered metabolic reactions fill gaps in the metabolic network maps, facilitating exploration of uncharted territories in cellular biochemistry.
    DOI:  https://doi.org/10.1038/s41467-025-60258-7
  22. Cancer Metastasis Rev. 2025 Jun 04. 44(2): 53
      Despite recent advances in targeted and immuno-therapy, metastasis still kills most cancer patients. Overcoming cancer metastasis requires a leap forward in understanding its molecular underpinnings to identify breakthrough therapeutic targets and strategies. A growing body of evidence suggests that specific subsets of cancer cells, which exhibit stem-like properties and are referred to as cancer stem cells (CSCs), are the primary drivers of cancer metastasis. How CSCs contribute to the multistep process of invasion and metastasis remains incompletely understood. Invadosomes are dynamic actin-based cellular protrusions that mediate cell invasion and pericellular proteolysis. Recent data have highlighted the highly proficient ability of CSCs to generate invadosomes, facilitating their local invasiveness, intravasation, extravasation, and metastatic colonization. This up-to-date and focused review describes the recent progress in characterizing invadosomes in embryonic cells during development and in CSCs during cancer metastasis. We summarize the molecular processes that regulate the invadosomes of CSCs. We discuss the molecules associated with the invadosome of CSCs and highlight their correlation with cancer metastasis in patients. We propose targeting the invadosomes of CSCs as a novel strategy to overcome cancer invasiveness and metastasis. This review highlights the emerging role of invadosomes in CSCs and provides a new perspective on pathobiology and cancer metastasis treatment.
    Keywords:  Cancer stem cell; Invadosome; Metastasis; Therapeutic target
    DOI:  https://doi.org/10.1007/s10555-025-10270-6
  23. Nat Protoc. 2025 Jun 03.
      Membrane fusion is central to cellular signaling and trafficking, requiring a detailed understanding of protein-lipid interactions. Studying these dynamic events in live cells presents challenges due to their complexity and heterogeneity. To address this, we developed a reductionist in vitro membrane model system that enables the controlled investigation of individual molecular components. This approach begins with a minimal membrane environment, with the opportunity for the stepwise addition of specific components to incrementally increase complexity achieving a level of experimental precision often unattainable in cellular studies. We developed suspended lipid membranes, a platform that uses pore-spanning lipid bilayers formed on microfabricated silicon chips with micrometer-sized holes. These membranes closely mimic native cellular architecture by maintaining aqueous compartments on both sides, providing a solvent-free, near-native environment with exceptional lateral diffusion properties. Their high stability makes them ideal for time-lapse imaging and dynamic process analysis using total internal reflection fluorescence and confocal microscopy. Here we present a detailed protocol for generating pore-spanning, planar suspended lipid membranes from native and synthetic reconstituted lipids using our silicon chip platform. Using SNARE proteins and molecular chaperones, we demonstrate the system's ability to capture ultrafast membrane fusion events. Additionally, we demonstrate single-molecule protein counting, protein dynamics analysis and single-vesicle fusion assays using fluorescently labeled proteins and vesicles. The ability to preserve native lipid asymmetry, biological composition and lateral diffusion makes this method a powerful tool for dissecting membrane fusion mechanisms and other membrane biological processes with unparalleled precision.
    DOI:  https://doi.org/10.1038/s41596-025-01192-2
  24. STAR Protoc. 2025 May 29. pii: S2666-1667(25)00215-1. [Epub ahead of print]6(2): 103809
      Atomic force microscopy (AFM) enables high-resolution mechanical characterization of soft materials at the nanoscale. It offers unique advantages over conventional mechanical testing methods by providing spatially resolved properties, requiring minimal sample preparation, and allowing measurements under controlled environmental conditions. This comprehensive guide provides a practical framework for conducting reproducible nanomechanical measurements on soft matter using AFM. Readers will learn how to select appropriate AFM modes, choose and calibrate suitable cantilevers, prepare samples, and optimize measurement parameters for soft materials. Four operational AFM modes are described: intermittent contact mode, nanomechanical imaging, force modulation, and force spectroscopy. We detail their principles, mechanisms, and trade-offs while offering practical advice for experiment execution, data analysis, and result reporting. This protocol seeks to guide researchers to execute consistent and comparable AFM measurements, bridge the gap between theoretical knowledge and practical implementation, and address key challenges in standardization and reproducibility within the field of soft matter nano-mechanics.
    Keywords:  Atomic Force Microscopy (AFM); Chemistry; Material sciences; Physics
    DOI:  https://doi.org/10.1016/j.xpro.2025.103809
  25. Lancet Gastroenterol Hepatol. 2025 May 16. pii: S2468-1253(25)00056-1. [Epub ahead of print]
    METAPAC trial investigators
       BACKGROUND: Earlier diagnosis of pancreatic ductal adenocarcinoma is key to improving overall survival in patients with this hard-to-treat cancer. We independently validated two previously identified plasma-based metabolic signatures for exclusion of pancreatic ductal adenocarcinoma in cohorts with an increased annual risk.
    METHODS: The METAPAC study was a prospective, multicentre, investigator-masked, enrichment design, phase 4 trial done in 23 centres in Germany. Patients with pancreatic lesions identified by diagnostic imaging that required further diagnostic assessment were recruited and followed up for 24 months. Targeted quantitative plasma metabolite analysis was done on a liquid chromatography-tandem mass spectrometry platform. The improved metabolic (i-Metabolic) signature consisted of 12 analytes plus carbohydrate antigen (CA) 19-9, and the minimalistic metabolic (m-Metabolic) signature consisted of four analytes plus CA 19-9. The primary endpoint of the study was the exclusion of pancreatic ductal adenocarcinoma with an 85% specificity and the highest possible diagnostic accuracy. All statistical analyses were done per protocol. This study is registered with the German Clinical Trials Register (DRKS00010866).
    FINDINGS: Between Sept 9, 2016, and April 8, 2022, 1370 patients with CT-identified pancreatic lesions necessitating further diagnostic assessment were screened, of whom 1129 patients (489 with pancreatic ductal adenocarcinoma, 640 controls) were included in the primary analysis (median age 67 years [IQR 58-75]; 556 [49%] female, 572 [51%] male). The control group consisted of high-risk individuals with acute pancreatitis (11 [1%] of 1129 participants), chronic pancreatitis (113 [10%]), intraductal papillary mucinous neoplasms (232 [21%]), cystic lesions other than intraductal papillary mucinous neoplasms (271 [24%]), and metastases of extrapancreatic origin (13 [1%]). The i-Metabolic signature detected pancreatic ductal adenocarcinoma with an area under the curve (AUC) of 0·846 (95% CI 0·842-0·849), specificity of 90·4% (89·8-91·1), sensitivity of 67·5% (66·9-68·0), and balanced accuracy of 80·5% (80·2-80·8), compared with CA 19-9 alone (AUC 0·799 [0·797-0·802], p<0·0001; specificity 79·1% [78·7-79·4]; sensitivity 81·8% [81·5-82·0]; balanced accuracy 80·6% [80·4-80·9]). The m-Metabolic signature detected pancreatic ductal adenocarcinoma with an AUC of 0·846 (95% CI 0·842-0·849; p<0·0001 vs CA 19-9 alone), specificity of 93·6% (93·1-94·0), sensitivity of 59·9% (59·3-60·4), and accuracy of 79·0% (78·8-79·2). In a population of 242 individuals with new-onset diabetes (three cases of incident pancreatic ductal adenocarcinoma), the m-Metabolic signature (without CA 19-9) significantly discriminated patients with pancreatic ductal adenocarcinoma from those without (p=0·038). AUC, specificity, and sensitivity remained constant after random bootstrapping for a prevalence of pancreatic ductal adenocarcinoma between 1% and 20%.
    INTERPRETATION: Two plasma-based metabolic signatures showed significant improvement in performance compared with CA 19-9 alone in excluding pancreatic ductal adenocarcinoma in a prospective real-world cohort. These findings could offer a surveillance tool in patients with an annual risk of pancreatic ductal adenocarcinoma of 1% to reduce unnecessary invasive procedures and facilitate earlier detection of resectable disease.
    FUNDING: Federal Ministry of Education and Research (BMBF, Germany).
    DOI:  https://doi.org/10.1016/S2468-1253(25)00056-1
  26. Nat Commun. 2025 Jun 05. 16(1): 5237
      Multivalent proteins can form membraneless condensates in cells by liquid-liquid phase separation, and significant efforts have been made to study their biochemical properties. Here, we demonstrate the emergent mechanics of a functional multivalent condensate reconstituted with six postsynaptic density proteins, using atomic-force-microscopy-based mesoscale rheology and quantitative fluorescence measurements. The measured relaxation modulus and protein mobility reveal that the majority (80%) of the proteins in the condensate are mobile and diffuse through a dynamically cross-linked network made of the remaining (20%) non-mobile scaffold proteins. This percolating structure gives rise to a two-mode mechanical relaxation with an initial exponential decay followed by a long-time power-law decay, which differs significantly from simple Maxwell fluids. The power-law rheology with an exponent α ≃ 0.5 is a hallmark of weak bonds' binding/unbinding dynamics in the multivalent protein network. The concurrent molecular and mechanical profiling thus provides a reliable readout for characterizing the mechanical state of protein condensates and investigating their physiological functions and associations with diseases.
    DOI:  https://doi.org/10.1038/s41467-025-60345-9
  27. Cancer Res. 2025 Jun 06.
      Normal tissues actively employ a phenomenon called cell competition to drive the elimination and replacement of less fit loser cells by fitter winner cells. This quality control mechanism promotes tissues health, by favouring the selective expansion of fitter cells. Indeed, through cell competition, many mutant cells are eliminated from tissues by fitter normal cells. However, some oncogenic mutations can turn cells into super-competitors that outcompete normal cells, promoting tumorigenic growth and metastasis. Several cellular stresses have been associated with the loser status such as oxidative stress, DNA damage responses, unfolded protein response and mitochondrial dysfunction. By affecting these pathways, metabolism and dietary choices can regulate cellular fitness and cell competition. However, how these pathways affect competitive interactions in vivo, during the early establishment of mutant clones, is relatively little understood. Recent work from Hemalatha and colleagues introduces real-time fluorescence ratio metric imaging of NAD(P)H and FAD, to investigate cellular redox status - live and over time, at single cell level - as cells compete in the mouse epidermis. Their work demonstrates that redox status changes dynamically during competition between cell carrying oncogenic mutations. It further shows that drugs that modulate mitochondrial metabolism and cellular redox are strong modulators of cell competition. The introduction of live redox imaging will prove a powerful tool to further dissect how metabolic states affect cell competition in normal physiology and in tumorigenesis.
    DOI:  https://doi.org/10.1158/0008-5472.CAN-25-2374
  28. Nature. 2025 Jun 04.
      Chimeric antigen receptor (CAR) natural killer (NK) cell immunotherapy offers a promising approach against cancer1-3. However, the molecular mechanisms that regulate CAR-NK cell activity remain unclear. Here we identify the transcription factor cyclic AMP response element modulator (CREM) as a crucial regulator of NK cell function. Transcriptomic analysis revealed a significant induction of CREM in CAR-NK cells during the peak of effector function after adoptive transfer in a tumour mouse model, and this peak coincided with signatures of both activation and dysfunction. We demonstrate that both CAR activation and interleukin-15 signalling rapidly induce CREM upregulation in NK cells. Functionally, CREM deletion enhances CAR-NK cell effector function both in vitro and in vivo and increases resistance to tumour-induced immunosuppression after rechallenge. Mechanistically, we establish that induction of CREM is mediated by the PKA-CREB signalling pathway, which can be activated by immunoreceptor tyrosine-based activation motif signalling downstream of CAR activation or by interleukin-15. Finally, our findings reveal that CREM exerts its regulatory functions through epigenetic reprogramming of CAR-NK cells. Our results provide support for CREM as a therapeutic target to enhance the antitumour efficacy of CAR-NK cells.
    DOI:  https://doi.org/10.1038/s41586-025-09087-8
  29. NPJ Biol Phys Mech. 2025 ;2(1): 16
      Mild traumatic brain injury (mTBI) is an acute injury with immediate and medium-term symptom presentation. However, our mechanistic understanding of mTBI and how mechanical loading of soft cellular tissues leads to injury is limited. The aim of this review is to introduce this interdisciplinary field to non-experts and provide an overview of our current understanding of how mechanical trauma contributes to cellular injury. Here, we compare the significance of various measures of mechanical loading including strain magnitude, strain rate, loading mode, and frequency, and their relative significance for cell and tissue injury in in vitro and ex vivo experimental models reported in the literature. Interestingly, while it is difficult to define a precise injury threshold value based on strain magnitude alone, cellular injury is commonly observed at strain rates of >0.1 s-1, higher than rates observed in many normal cell functions (< 0.01 s-1). We explore the role of the plasma membrane, cytoskeleton, and specialized structures in maintaining cell integrity during traumatic injury.
    Keywords:  Biological techniques; Biophysics; Diseases
    DOI:  https://doi.org/10.1038/s44341-025-00020-8
  30. J Org Chem. 2025 Jun 02.
      Biphasic systems are ubiquitous across modern chemistry, yet direct and quantitative analysis of solute distribution across phases remains experimentally nontrivial. While many chemists rely on qualitative intuition to reason about partitioning, confirming these assumptions typically requires time-consuming sample manipulation. We demonstrate that slice-selective NMR spectroscopy─a straightforward technique available on modern NMR instruments─can be used to directly measure spatially resolved concentrations. This allows users to quantify how species partition between immiscible phases without disrupting the equilibrium, offering a powerful and broadly accessible tool for reaction monitoring, mechanistic study, and process troubleshooting. We demonstrate the application of this technique to measure logP values, monitor phase-transfer catalysis, study biphasic reaction kinetics, and quantify the influence of counterion identity and ionic strength on phase distribution. In each example, slice-selective NMR enables an understanding of underlying chemical behavior that would otherwise be difficult to capture. We aim to highlight this technique not as a niche spectroscopic curiosity but as a general-purpose tool that can be readily adopted by synthetic and process chemists. This work serves as both a tutorial introduction and a practical demonstration of how spatially selective NMR can demystify the behavior of biphasic systems and bring quantitative rigor to problems often approached qualitatively.
    DOI:  https://doi.org/10.1021/acs.joc.5c00717
  31. bioRxiv. 2025 May 18. pii: 2025.05.15.654206. [Epub ahead of print]
      Cold stress elicits dynamic remodeling of the mitochondrial lipidome in brown adipose tissue (BAT), marked by an increase in arachidonoyl-phosphatidylethanolamine (AA-PE). However, the function of membrane lipid rewiring in thermoregulatory physiology has been a longstanding mystery. Here, we identify LPCAT3 as a cold-regulated O-acyltransferase driving the highly selective accrual of AA-PE in BAT mitochondria. Lipid-based proteomics, molecular dynamics simulations, and bioenergetic analyses reveal that AA-PE partitions at the COX4I1 interface of the Cytochrome c oxidase complex, enhancing electron transport chain (ETC) efficiency. Accordingly, fat-specific Lpcat3 -knockout mice have defects in respiratory-dependent BAT thermogenesis and cold tolerance, despite intact β-adrenergic signaling and UCP1 function. Under cold acclimation, Lpcat3 -/- BAT exhibits ETC dysfunction and activation of the integrated stress-response. Thus, our study illuminates a cold-regulated lipid-protein interaction as a gating factor in UCP1-dependent thermogenesis.
    DOI:  https://doi.org/10.1101/2025.05.15.654206
  32. J Biol Chem. 2025 May 29. pii: S0021-9258(25)02161-1. [Epub ahead of print] 110311
      Despite the well described role of non-cellular components of the tumor microenvironment (TME) in regulating tumor growth, the molecular events dictating expression and biological functions of key components of the TME remain elusive. Here, using pancreatic cancer (PC) models, we describe a novel mechanism through which the zinc finger transcription factor GLI2 in cancer associated fibroblasts (CAFs) induces expression of COL1A1, which is a major component of Type I Collagen, the most abundant collagen variant in the tumor milieu. Bulk and single nuclei RNA-Seq showed that GLI2 expression in CAF strongly correlates with COL1A1 expression levels, fibrosis, and CAF activation. ChIP-qPCR and expression studies of the PC matrisome identified COL1A1 as the direct target of GLI2 in CAFs. We also provide evidence that GLI2 is an effector that mediates COL1A1 induction by transforming growth factor β1 (TGFβ1). RNA-Seq analysis of PC cells treated with Type I Collagen revealed enrichment of chemotherapeutic gene expression profiles, which includes irinotecan resistance signature. Viability studies confirmed that Type I Collagen promotes irinotecan resistance in PC cells. Altogether, our results uncover a novel role for the TGFβ1-GLI2 axis within CAFs to modulate Type I Collagen expression and promote chemoresistance in PC cells. Together, our findings help increase the understanding of the complex molecular network operating in the TME.
    Keywords:  GLI2; collagen; fibroblast; pancreatic cancer; tumor microenvironment
    DOI:  https://doi.org/10.1016/j.jbc.2025.110311
  33. Nat Commun. 2025 Jun 05. 16(1): 5231
      Stress stimulation-mediated liquid-liquid phase separation is a key activity in living organisms, but its biophysical characteristics are poorly understood. Here, we report a UV-light stress stimulation behaviour in a binary community of synthetic protocells of condensates and proteinosomes, showing that condensates could behave like Condensate Pumps to enable a stepwise controlled transmembrane mass transfer regardless of the permeability barrier of proteinosomes. The stimulation mechanism of interfacial tension-induced proteinosome deformation and transient high osmotic pressure arisen by the dissociation of condensate is proposed. Accordingly, under UV-light stress stimulation, unexpected characteristics could be triggered by transmembrane pumping oversized biomacromolecules into proteinosomes including liquid-liquid reentrant phase separation, DNA unwinding, and protein synthesis. Therefore, our results not only reveal unique physical principles and potential characteristics of macromolecular assemblies at droplet-membrane interface but also highlight a pathway for transmembrane transport of biomacromolecules which is anticipated to serve as a powerful technique to inducing higher-order behaviour in synthetic protocells community.
    DOI:  https://doi.org/10.1038/s41467-025-60541-7
  34. ACS Sens. 2025 Jun 06.
      Microfluidic cell separation by dielectrophoresis, based on biophysical and electrical physiology metrics, is often optimized using on-chip fluorescence microscopy or off-chip flow cytometry of the separated fractions. However, these techniques require fluorescent reporters or stained samples that operate as end point assays, preventing the separated cell fractions from being utilized for longitudinal or transplantation studies. Single-cell impedance cytometry has a small footprint for facile integration toward label-free quantification of the separated fractions based on cell size, viability, and biophysical metrics. However, this is limited by low impedance signal-to-noise ratios in the low-conductivity media optimal for dielectrophoretic separation and by irreversible dielectrophoretic cell capture on impedance acquisition electrodes, while its single-cell resolution ability is limited by the high channel depths used to enhance sample throughput. Herein, using viscoelastic flows for elasto-inertial cell focusing over the channel depth, the throughput of dielectrophoretic separation is maintained, and bubble formation is avoided, while the downstream voltage for impedance cytometry can be maximized without irreversible cell capture to enhance impedance sensitivity in the dielectrophoretic separation media. This multichannel cytometry capability is integrated for automated optimization of the dielectrophoretic enrichment of live circulating tumor cells released into the suspension of pancreatic cancer cell cultures, using impedance metrics to monitor the separated fractions for feedback toward the selection of live cells within specific size ranges and with minimized transmembrane voltage-induced cell damage.
    Keywords:  automation; circulating tumor cells; dielectrophoresis; impedance Cytometry; microfluidics; single-cell analysis
    DOI:  https://doi.org/10.1021/acssensors.5c00192