Biology subjects

Lee, J. K.

Publications and source records attributed to Lee, J. K..

At least 19 recordsLinked to original sources

bioRxiv2026

Vocal fold injury alters acoustic phenotype despite preserved vocal activity

Vocal communication depends on the biomechanical integrity of the vocal folds, yet how impairment alters vocal behavior remains poorly understood. We used the California mouse (Peromyscus californicus), a highly vocal species that produces songs through airflow-induced vocal fold vibration, to experimentally examine how defined vocal fold injury alters vocal production. Longitudinal recordings were combined with histological assessment to track changes in vocal function after impairment. Injury produced acute edema of the lamina propria and thyroarytenoid muscle. Despite stable levels of vocal activity, the vocal phenotype shifted characteristically following injury. Thus, vocal behavior can persist despite substantial alteration of the biomechanical tissues underlying sound production, demonstrating that maintenance of behavioral output does not necessarily indicate preservation of the physiological system that generates it. More broadly, these findings establish an experimental link between vocal fold injury and vocal phenotype and provide a framework for understanding how tissue damage and remodeling can reshape acoustic communication even when vocal behavior is maintained. Because repeated vocalization can impose substantial mechanical demands on vocal folds, this framework may have broad relevance for understanding how vocal use, injury, and tissue remodeling interact to shape vocal communication.

physiology↗

bioRxiv2026

Water oxidation-driven histidine dioxidation enables probe-free proximity labeling

Photocatalytic proximity labeling (photo-PL) has emerged as a powerful tool for spatial proteomics in subcellular compartments. However, many photo-PL toolboxes rely on singlet oxygen (1O2) to generate highly unstable endoperoxide intermediates that must be trapped immediately by high concentrations of exogenous probes. This constraint can bias spatial proteome coverage, particularly in dynamic and heterogeneous compartments such as endosomes and exosomes, where probe accessibility is intrinsically nonuniform. Here, we develop IDM, an organic photocatalyst that generates hydroxyl (*OH) and superoxide (O *-) radicals via water oxidation instead of 1O , enabling a synergistic dual-radical mechanism for probe-free proximal protein mapping in live cells (PF-Map). The resulting radicals dioxidize proximal histidine residues into a persistent dioxidized state (His-2O) that is thermodynamically stabilized as lactam tautomers, which remain electrophilic and chemically addressable after cell lysis. By decoupling histidine oxidation from live{square}cell probe capture, PF{square}Map can minimize spatial bias arising from heterogeneous probe distribution. Applying PF{square}Map to intracellular vesicle trafficking, we find that both PF{square}Map and a probe{square}dependent workflow (PD{square}Map) robustly identify exosome markers, whereas PF{square}Map additionally reveals a hidden vesicle trafficking-related subproteome that PD-Map underestimated. Together, we establish a minimally biased photocatalytic strategy for spatial protein mapping in complex biological systems. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=111 SRC="FIGDIR/small/731337v1_ufig1.gif" ALT="Figure 1"> View larger version (35K): org.highwire.dtl.DTLVardef@13da43dorg.highwire.dtl.DTLVardef@1e9cb7corg.highwire.dtl.DTLVardef@2e7557org.highwire.dtl.DTLVardef@19e16c_HPS_FORMAT_FIGEXP M_FIG C_FIG

biochemistry↗

bioRxiv2026

Augmenting the Bayesian Brain with learned and reusable world-model components for flexible cognition

The Bayesian Brain hypothesis assumes that cognition relies on internal generative models of the world, yet existing implementations remain constrained by pre-specified, task-specific generative structures and computationally heavy iterative inference schemes. Here, we introduce modular neural state-space models as a scalable realization of the Bayesian Brain, replacing fixed generative structures and pre-specified inference rules with learned world-model components and amortized neural updates. This framework preserves the core commitment to explaining observations through hidden causes while making inference learned and reusable rather than pre-specified and task-specific. Our modular implementation of these models affords learned components to be seamlessly recombined and stacked across superficially different tasks that share similar latent dynamics. Such computational reuse supports zero-shot generalization and predicts selective correlations of inference parameters between tasks. We confirm these key predictions in human behavior, identifying learned and reusable world-model components as a candidate computational principle for flexible cognition.

neuroscience↗

bioRxiv2026

Structure-guided deimmunisation of Fel d 1 suppresses allergic effector functions ex vivo

Allergic responses to the major cat allergen Fel d 1 are primarily driven by immunoglobulin E (IgE) cross-linking on effector cells. While conventional allergen-specific immunotherapies often rely on whole-allergen extracts with inherent risks of systemic reactions, rationally engineered variants offer a safer path to desensitisation. We present a structure-guided strategy to deimmunise Fel d 1 by selectively disrupting crucial IgE-binding interfaces. Using a computational pipeline integrating structural mapping and immunoinformatics, we designed 30 single-point mutations targeting immunodominant regions. Ex vivo functional evaluation using sera from cat-allergic individuals demonstrated that specific variants, notably K29G in Chain 1, exhibited significantly diminished IgE reactivity and profoundly suppressed basophil activation. Furthermore, CRISPR/Cas9-mediated introduction of the K29G mutation into feline fibroblasts confirmed that the substitution preserves fundamental cellular proliferation. This work demonstrates that targeted computational design can not only yield superior candidates for safer immunotherapeutics but also establish a viable foundation for generating genetically edited, hypoallergenic cats.

immunology↗

bioRxiv2026

Systematic mapping of emergent transcriptional states in interacting single-cell dyads by Cell-Cell-seq

Cell-cell interactions drive rapid and heterogeneous changes in gene expression, yet most transcriptomic methods either dissociate cells, losing pair identity and interaction timing, or infer communication indirectly from ligand-receptor co-expression. Here we present Cell-Cell-seq, a scalable workflow for profiling defined cell pairs ("dyads") with single-cell transcriptomic resolution. Cell-Cell-seq uses cavity-containing hydrogel microparticles (Nanovials) to confine two cells, synchronize contact onset, protect fragile conjugates during handling and sorting, and interface directly with droplet-based RNA sequencing. Using antigen-matched prostate tumor cells and engineered T cells as a model system, Cell-Cell-seq captured thousands of tumor-T cell dyads and revealed broad functional and transcriptional heterogeneity across interactions. Dyads unmasked transient activation programs that were obscured in standard well-plate co-culture, consistent with asynchronous contact in bulk assays. To distinguish interaction-induced programs from the composite nature of dyad transcriptomes, we developed a pseudo-mixing framework that generates in silico pseudo-dyads to construct an empirical null distribution under "no interaction," enabling statistically robust identification of emergent genes and partner-resolved attribution of responses. Dyad-resolved analysis further revealed coordinated cross-cell programs, including coupled chemokine expression consistent with bidirectional paracrine signaling and inverse coupling between tumor immunoregulatory programs and T cell activation. Finally, we introduce ccRepair to correct compositional dilution in mixed transcriptomes, improving interpretability while preserving genuine cross-cell coordination. Together, Cell-Cell-seq provides a generalizable platform for dissecting immune synapse biology and mapping interaction-dependent programs across heterogeneous cell populations, with applications in profiling tumor-immune communication and functionally screening immunotherapies.

bioengineering↗

bioRxiv2025

Armoring STEAP1 CAR T cells with IL-18 potentiates antitumor activity in Ewing sarcoma

BackgroundEwing sarcoma (EwS) is a highly aggressive cancer driven by the EWS::FLI1 fusion oncoprotein affecting children, adolescents, and young adults. Six transmembrane epithelial antigen 1 (STEAP1) is a cell surface antigen transcriptionally controlled by EWS::FLI1 that is broadly expressed in EwS, positioning it as a rational immunotherapy target. However, translating CAR T therapy to solid tumors requires overcoming barriers to potency while maintaining safety. MethodsAnalyses of transcriptome and proteome data were performed to evaluate the effects of EWS::FLI1 perturbation on STEAP1 expression at the transcript and protein levels in EwS models. STEAP1 expression was validated in EwS patient tissues by immunohistochemistry. Second-generation STEAP1-BB{zeta} CAR T cells were tested in orthotopic and disseminated EwS xenograft models. To enhance antitumor activity, an IL-18-armored STEAP1 CAR was engineered. Dose-dependent therapeutic efficacy and safety were evaluated through measurement of tumor burden, survival, and observation for gross toxicities. ResultsSTEAP1 was expressed in [~]97% of primary EwS tumors and directly associated with EWS::FLI1 fusion protein expression in EwS cell lines. In orthotopic EwS models, STEAP1 CAR T cells induced complete tumor regression at 5 x 106 cells. In disseminated disease models, responses were dose-dependent with no evidence of antigen loss. Notably, IL-18 armored STEAP1 CAR T cells achieved complete responses in [~]80% of mice at a reduced dose of 106 cells without overt toxicity. ConclusionsThese data establish STEAP1 as a clinically relevant and highly expressed target in EwS and demonstrate that IL-18 armoring significantly improves CAR T cell efficacy by enhancing potency evident through antitumor activity at reduced cell dose. STEAP1 CAR T cells are currently under evaluation in a first-in-human phase 1/2 dose-escalation clinical trial for metastatic castration-resistant prostate cancer (NCT06236139) and these studies support future clinical translation of STEAP1 CAR T cell therapy for relapsed/refractory EwS.

cancer biology↗

bioRxiv2025

A dual information seeking strategy improves imprecise human inferences outside the explore-exploit tradeoff

Everyday decisions aim not only to earn rewards but also to learn about the world. Across three studies (total N = 702), we examined how people gather epistemic information stripped of rewarding value, and compared their strategy with reward seeking in otherwise matched conditions. Computational modeling of human behavior revealed a two-stage information-seeking policy, where participants first repeatedly sample each novel option in turn to test provisional hypotheses, a process we call streaking, before transitioning to uncertainty-guided exploration. While artificial neural networks trained to optimize inference accuracy acquired uncertainty-guided exploration but not early streaking, this two-stage policy improves human inference accuracy under noisy belief updating. Streaking and uncertainty-guided exploration tend to be co-expressed in the same individuals but map onto distinct psychological traits. Together, these results offer a novel account of human information seeking, clarifying its motives and benefits in epistemic contexts beyond the reward-centric explore-exploit tradeoff.

neuroscience↗

bioRxiv2025

FDA-approved drug repurposing in zebrafish identifies thyroid hormone and other compounds as potential antithrombotics

Venous thromboembolism (VTE) is a highly prevalent medical condition with limited therapeutic options and an incomplete understanding of its acquired and inherited subtypes. The zebrafish is a model with the benefits of external development, fecundity, optical transparency, and hemostasis that demonstrates conservation with mammals. We utilized zebrafish as a phenotypic screening tool to identify novel therapeutic options for preventing VTE. A library of FDA-approved compounds was screened for suppression of acquired (elevated estrogen) and spontaneous (protein C deficiency) thrombosis. We found that thyroid hormone, receptor tyrosine kinase (RTK) inhibitors, and proton-pump inhibitors (PPIs) effectively modulated levels of thrombosis, particularly in the estrogen-induced model. These also showed a more favorable hemostatic profile than standard therapies, suggesting alternative mechanisms. Genome editing of thyroid hormone receptor proved that thyroid hormone action is on target. A retrospective electronic health record (EHR) analysis found that thyroid-hormone prescriptions in hormonal contraceptive users correlated with a higher VTE risk, potentially limiting direct repurposing but highlighting thyroid signaling as a pathway involved in estrogen-induced thrombosis. Together, these data identify several drug classes that can be tailored to specific subtypes of VTE and help elucidate distinct pathways driving thrombosis.

pharmacology and toxicology↗

bioRxiv2025

Genotoxic antibody-drug conjugates combined with Bcl-xL inhibitors enhance therapeutic efficacy in metastatic castration-resistant prostate cancer

Metastatic castration-resistant prostate cancer (mCRPC) is an aggressive subtype of prostate cancer (PC) without curative treatments. Antibody-drug conjugates (ADCs) emerged as promising cancer therapeutics that selectively deliver cytotoxic agents (payloads) to the tumors. Although ADCs have been successfully applied in the treatment of hematological and solid tumors, ADC monotherapy has not demonstrated durable responses in mCRPC and the mechanisms of PC resistance to ADCs have not been thoroughly investigated. Our study aimed to improve ADC efficacy using a new integrated approach for custom ADC design and multiplexing. To nominate rational combinations of ADC targets and ADC payloads, we (1) examined protein co-expression of three clinically relevant surface antigens-- B7 homolog 3 (B7-H3), prostate specific membrane antigen (PSMA), and six-transmembrane epithelial antigen of prostate-1 (STEAP1)--in a series of human mCRPC samples and (2) screened established ADC payloads and their combinations in mCRPC cell lines with different phenotypes. We identified synergistic interactions between DNA-damaging payloads and Bcl-xL inhibitor A-1331852 as well as their coordinated induction of the intrinsic apoptosis pathway. The functional relevance of isolated p53 loss and impaired PC responses to three genotoxic ADCs (B7-H3-seco-DUBA, PSMA-SG3249, and STEAP1-DXd) and their combinations with A-1331852 was established using genetic knockout models. Lastly, we found enhanced in vivo antitumor activity in mCRPC by combining the clinically relevant agents B7-H3-seco-DUBA (vobramitamab duocarmazine) and A-1331852. Collectively, our findings provide rationale for the development of ADC therapies combining genotoxic payloads with Bcl-xL inhibitors for mCRPC. SignificanceB7-H3, PSMA, and STEAP1 targeted ADC therapies combining genotoxic payloads with Bcl-xL inhibitors induce p53-dependant apoptotic cell death in mCRPC, providing a clinically viable strategy for the treatment of advanced prostate cancer.

cancer biology↗

bioRxiv2025

Gluconeogenesis in Aqueous Microdroplets: Non-Enzymatic Generation of Glucose

Glucose is a central metabolite of living organisms, serving as the primary energy substrate produced predominantly by photosynthetic organisms. Under conditions of limited external glucose supply, organisms activate gluconeogenesis, an endogenous biosynthetic pathway that sustains essential glucose levels. Yet the mechanism of glucose generation, before the advent of photosynthesis and complex enzymatic systems, remains elusive. Recently, microdroplet chemistry has emerged as a novel approach for catalyst-free organic synthesis. Here, we report the non-enzymatic formation of glucose from a simpler organic precursor, pyruvate, in aqueous microdroplets without the aid of organic or inorganic catalysts. Our results show that glucose is generated via a reaction pathway analogous to canonical gluconeogenesis, proceeding through key intermediates including oxaloacetate, glycerate, and glyceraldehyde. Furthermore, thermodynamic analysis indicates that the free energy change associated with glucose formation is overcome in aqueous microdroplets at room temperature, without the need for external energy input or enzymatic catalysis. These findings indicate that aqueous microdroplets can non-enzymatically convert C3 compound, pyruvate, into the C6 sugar glucose, offering a plausible abiotic route for anabolic carbon transformations during abiogenesis.

evolutionary biology↗

bioRxiv2025

Molecular pathology of acute spinal cord injury in middle-aged mice

The median age of sustaining a spinal cord injury has steadily increased from 29 to 43 over the last several decades. Although more pre-clinical studies in aged rodents are being done to address this shift in demographics, there have not been comprehensive transcriptomic studies investigating SCI pathobiology in middle-aged mice. To address this gap in knowledge, we compared behavioral, histopathological, and transcriptional outcomes in young (2-4 months-old) and middle-aged (10-12 months-old) mice. In contrast to previous studies, open field tests showed no differences in locomotor recovery between the young and middle-aged mice over a one-month period. The injury site also demonstrated similar histopathology in terms of lesion size, and numbers of macrophages and fibroblasts. Acutely after injury, proliferation of macrophages, fibroblasts, and astrocytes were also similar between the two age groups. In addition, spatial transcriptomics showed similar transcriptionally defined regions around the injury site at 3 days post injury. However, single cell RNA-sequencing of the cells at the injury site and surrounding spared tissue showed differences in select cell subpopulations. Taken together, our results indicate that although young and middle-aged mice display similar locomotor recovery and histopathology after SCI, changes in cell subpopulations may underly a decline in repair mechanisms that are manifested after this age.

neuroscience↗

bioRxiv2025

Solid tumor CAR T cells engineered with fusion proteins targeting PDL1 for localized IL-12 delivery

CAR T cell efficacy in solid tumors is limited due in part to the immunosuppressive TME. To improve anti-tumor responses, we hypothesized that enabling CAR T cells to secrete bifunctional fusion proteins consisting of a cytokine modifier (e.g., TGF{beta}trap, IL15, or IL12) combined with an immune checkpoint inhibitor (e.g., PDL1) will provide tumor localized immunomodulation to improve CAR T cell functionality. To that end, we engineered CAR T cells to secrete TGF{beta}trap, IL15, or IL12 molecules fused to PDL1 scFv, and assessed in vitro functionality and in vivo safety and efficacy in prostate and ovarian cancer models. CAR T cells engineered with PDL1-IL12 were superior in safety and efficacy compared to CAR T cells alone and to those engineered with PDL1 fused with TGF{beta}trap or IL15. Further, PDL1-IL12 engineered CAR T cells improved T cell trafficking and tumor infiltration, localized IFN{gamma} production, TME modulation, and anti-tumor responses, with reduced systemic inflammation-associated toxicities. We believe our PDL1-IL12 engineering strategy presents an opportunity to improve CAR T cell clinical efficacy and safety across multiple solid tumor types.

bioengineering↗

bioRxiv2025

Targeting the ITGA3B1 Complex with A Novel Antibody-Drug Conjugate for Metastatic Bladder Cancer

Advances in antibody (Ab) phage display technologies have enabled the development of targeted therapies with enhanced precision and clinical efficacy. Here, we report a phenotypic screening strategy combining target-unbiased live-cell biopanning with in situ chemical crosslinking and mass spectrometry to identify internalizing antibodies and their cognate antigens--key features for effective antibody-drug conjugate (ADC) development. Using this approach, we identified the 2E7 antibody, which exhibits rapid internalization and high specificity for the integrin 3{beta}1 (ITGA3B1) heterodimer, a complex overexpressed in multiple solid tumors, including bladder cancer. We generated an ITGA3B1-targeted ADC by conjugating monomethyl auristatin E (MMAE) to the 2E7 antibody, enabling selective delivery of cytotoxic payloads to ITGA3B1-positive cells. In preclinical bladder cancer models, this ADC demonstrated potent and dose-dependent antitumor efficacy, with significant tumor regression and improved survival. Our findings establish a framework for target discovery using live-cell phenotypic screening and position ITGA3B1 as a promising therapeutic target for ADC-based treatment of advanced bladder cancer. TeaserLive-cell biopanning uncovers ITGA3B1 as a target for ADC development in metastatic bladder cancer.

synthetic biology↗

bioRxiv2025

Collagen-binding IL-12 expressing STEAP1 CAR-T cells reduce toxicity and eradicate mouse prostate cancer in combination with checkpoint inhibitors

Immunosuppressive microenvironments, the lack of immune infiltration, and antigen heterogeneity pose significant challenges for chimeric antigen receptor (CAR)-T cell therapies to tackle solid tumors. CAR-T cells were armed with immunostimulatory payloads, such as interleukin-12 (IL-12), to overcome this issue, but faced intolerable toxicity during clinical development. Here, we show that collagen-binding domain-fused IL-12 (CBD-IL-12) was retained within syngeneic murine prostate tumors, after secretion from CAR-T cells targeting human six transmembrane epithelial antigen of the prostate 1 (STEAP1). This led to equivalently high intratumoral interferon-{gamma} levels without hepatotoxicity and infiltration of T cells into non-target organs, compared with unmodified IL-12. Both innate and adaptive immune compartments were dramatically activated and recognized diverse tumor antigens after CBD-IL-12 CAR-T cell treatment. Combination immunotherapy of CBD-IL-12 CAR-T cells and immune checkpoint inhibitors eradicated large tumors in an established prostate cancer model, without pre-conditioning chemotherapy. The therapy generated anti-tumor immunological memory. CBD-fusion to potent yet toxic payloads of CAR-T therapy may remove obstacles to their clinical translation towards elimination of solid tumors.

bioengineering↗

bioRxiv2025

Defined cellular reprogramming of androgen receptor-active prostate cancer to neuroendocrine prostate cancer

Neuroendocrine prostate cancer (NEPC) arises primarily through neuroendocrine transdifferentiation (NEtD) as an adaptive mechanism of therapeutic resistance. Models to define the functional effects of putative drivers of this process on androgen receptor (AR) signaling and NE cancer lineage programs are lacking. We adapted a genetically defined strategy from the field of cellular reprogramming to directly convert AR-active prostate cancer (ARPC) to AR-independent NEPC using candidate factors. We delineated critical roles of the pioneer factors ASCL1 and NeuroD1 in NEtD and uncovered their abilities to silence AR expression and signaling by remodeling chromatin at the somatically acquired AR enhancer and global AR binding sites with enhancer activity. We also elucidated the dynamic temporal changes in the transcriptomic and epigenomic landscapes of cells undergoing acute lineage conversion from ARPC to NEPC which should inform future therapeutic development. Further, we distinguished the activities of ASCL1 and NeuroD1 from the inactivation of RE-1 silencing transcription factor (REST), a master suppressor of a major neuronal gene program, in establishing a NEPC lineage state and in modulating the expression of genes associated with major histocompatibility complex class I (MHC I) antigen processing and presentation. These findings provide important, clinically relevant insights into the biological processes driving NEtD of prostate cancer.

cancer biology↗

bioRxiv2024

OrthologAL: A Shiny application for quality-aware humanization of non-human pre-clinical high-dimensional gene expression data

Single-cell and spatial transcriptomics provide unprecedented insight into the inner workings of disease. Pharmacotranscriptomic approaches are powerful tools that leverage gene expression data for drug repurposing and treatment discovery in many diseases. Multiple databases attempt to connect human cellular transcriptional responses to small molecules for use in transcriptome-based drug discovery efforts. However, pre-clinical research often requires in vivo experiments in non-human species, which makes capitalizing on such valuable resources difficult. To facilitate the application of pharmacotranscriptomic databases to pre-clinical research models and to facilitate human orthologous conversion of non-human transcriptomes, we introduce OrthologAL. OrthologAL leverages the BioMart database to access different gene sets from Ensembl, facilitating the interaction between these servers without needing user-generated code. Researchers can input their single-cell or other high-dimensional gene expression data from any species, and OrthologAL will output a human ortholog-converted dataset for download and use. To demonstrate the utility of this application, we characterized orthologous conversion in single-cell, single-nuclei, and spatial transcriptomic data derived from common pre-clinical models, including patient-derived orthotopic xenografts of medulloblastoma, and mouse and rat models of spinal cord injury. We show that OrthologAL can convert these data types efficiently to that of corresponding orthologs while preserving the dimensional architecture of the original non-human expression data. OrthologAL will be broadly useful for applying pre-clinical, high-dimensional transcriptomics data in functional small molecule predictions using existing human-annotated databases.

bioinformatics↗

bioRxiv2024

Hormone-induced thrombosis is mediated through non-canonical fibrin(ogen) aggregation and a novel estrogen target in zebrafish

Venous thrombosis is a well-known complication of estrogen exposure, with nearly every woman at risk across her lifetime through contraception, pregnancy, or hormone therapy. Although estrogens alter expression of coagulation factors, the mechanisms that mediate estrogen-induced thrombosis are poorly understood, partially due to the absence of an animal model. Identification of these mediators is central to understanding of hormone-induced pathophysiology, could ascertain patients at higher risk for thrombosis, and pinpoint future therapeutic targets. The zebrafish is characterized by external development, high fecundity, optical transparency, and hemostasis is highly conserved with humans. Through a transgenic line that generates GFP-tagged fibrinogen, we show rapid onset of thrombosis after exposure to various estrogens, but not progestins or testosterone. Thrombi are localized to the venous system with evidence for clot contraction. Thrombosis is only partially impeded by anticoagulants, occurs in the absence of factor VII, factor X, and prothrombin, but is dependent on tissue factor and fibrin(ogen). Finally, targeting of all known estrogen receptors does not eliminate thrombosis. The inability to completely inhibit thrombosis through genetic/pharmacologic anticoagulation or estrogen receptor disruption suggests mechanisms different from canonical coagulation/thrombosis. These studies suggest that estrogen-induced thrombosis is a unique entity distinct from other forms of venous thrombosis.

pathology↗

bioRxiv2024

Development of a Multifunctional Extracorporeal Life Support (ECLS) System for Lung and Kidney Support: The Pneuma-K ECLS System

Multi-organ failure (MOF), particularly in the coexistence of acute kidney injury (AKI) and acute lung injury (ALI), presents a significant challenge in intensive care units (ICU) and is associated with exceedingly high mortality rates. Respiratory and renal failures are frequently managed by extracorporeal membrane oxygenation (ECMO) and continuous renal replacement therapy (CRRT), respectively. However, employing these therapies using separate devices requires specialized facilities, adds to complexity, and increases the risks of clotting due to the extensive artificial surface areas involved. Therefore, an integrated device capable of providing simultaneous respiratory and renal support is essential. This paper introduces the Pneuma-K ECLS system, which incorporates a multifunctional detoxifying filter (MDF) capable of performing gas exchange and renal replacement in a single cartridge. Ex-vivo blood tests confirmed the ability of the MDF to oxygenate blood, remove carbon dioxide, and eliminate uremic toxins. In addition, animal experiments demonstrated the considerable clinical potential of this novel integrated extracorporeal life support approach. Integrating respiratory and renal support into a singular device could mitigate risks, conserve resources, and enhance the survival rates of critically ill patients suffering from concurrent lung and kidney failure.

bioengineering↗