Biology subjects

Cook, A.

Publications and source records attributed to Cook, A..

At least 19 recordsLinked to original sources

bioRxiv2026

Nuclear Myosin VI stabilises Ku-associated DNA ends during non-homologous end joining

DNA double-strand breaks (DSBs) require rapid signalling and physical stabilisation of broken DNA ends to preserve genome integrity. Here, we identify myosin VI (MVI) as an ATM-regulated component of the DSB response. DNA damage induces rapid nuclear accumulation and nanoscale reorganisation of MVI across multiple cell models, in an ATM-dependent manner. Pharmacological or genetic perturbation of MVI attenuates {gamma}H2AX signalling and disrupts Ku80 organisation, while DNA damage persists. This leads to increased sensitivity to cisplatin and bleomycin. Super-resolution imaging reveals spatial association of MVI with Ku80-containing repair structures, implicating MVI in non-homologous end joining (NHEJ). In a minimal reconstituted system, MVI and actin enhance the proximity of Ku70/80-bound DNA ends. Together, our findings identify MVI as a regulator of DSB repair that links ATM signalling to Ku-associated DNA-end stabilisation and suggest that targeting MVI may sensitise tumour cells to genotoxic therapy.

cancer biology↗

bioRxiv2026

Complementary Single-Cell Microflow HILIC and Ion Pair LC-MS Reveal Bystander Metabolic Effects in a Macrophage Model of Tuberculosis

Single-cell metabolomics remains analytically challenging due to the low abundance and chemical diversity of metabolites in individual cells. We have developed complementary microflow HILIC and ion pair LC-MS methods to expand metabolite coverage in single macrophages. Ion pair LC-MS was applied to single cells for the first time, enabling retention of highly polar and ionic metabolites that elute early under conventional reversed-phase conditions. Across Mycobacterium bovis BCG infected, uninfected bystander, and control unexposed THP-1 macrophages, both microflow methods detected significantly more features than a previously reported analytical-flow HILIC method. The two microflow methods provided complementary chemical space, together yielding 633 unique named metabolites with MS2 spectra. This depth enabled pathway-level interpretation at single-cell resolution, revealing infection-associated changes in purine-, arginine-, glutathione-, and one-carbon folate-associated metabolism. Metabolite-level interrogation indicated shared purine and amino acid changes in both infected and neighbouring macrophages, while revealing a distinct bystander phenotype characterised by elevated glycine and heterogeneous ATP levels. Finally, we demonstrate sequential IP and HILIC analysis of the same single cell, establishing a route toward maximal coverage from individual cells. These results position microflow HILIC and IP LC-MS as powerful, orthogonal strategies for advancing single-cell metabolomics and unveiling heterogeneity within complex biological microenvironments. Table of Contents O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=78 SRC="FIGDIR/small/733771v1_ufig1.gif" ALT="Figure 1"> View larger version (24K): org.highwire.dtl.DTLVardef@103ecfforg.highwire.dtl.DTLVardef@bc6734org.highwire.dtl.DTLVardef@11b96aforg.highwire.dtl.DTLVardef@41edfe_HPS_FORMAT_FIGEXP M_FIG Figure made in BioRender. C_FIG

microbiology↗

bioRxiv2026

Nanoflow ion-pairing LC-MS for ultra-low-input polar metabolomics and isotope tracing

Low-input and single-cell metabolomics remain constrained by the poor retention of polar metabolites in conventional reversed-phase nanoflow LC-MS workflows. Here, we establish nanoflow tributylamine (TBA) ion-pairing LC-MS as a platform for ultra-low-input polar metabolomics and stable isotope tracing. By adapting an analytical-flow TBA ion-pairing method to the nanoflow scale, this workflow extends the sensitivity and inline concentration advantages of nanoflow chromatography to charged metabolites involved in central carbon metabolism. Using mouse liver metabolite extracts, we show that the nanoflow method preserves chromatographic retention and separation of chemically diverse metabolite classes, including adenine nucleotides, nucleotide cofactors, TCA cycle intermediates, acyl-CoAs, and bile acid isomers. Despite loading 20-fold less tissue-equivalent material on column, nanoflow LC-MS produced higher signal intensity than the analytical-flow method for many metabolites. Across representative compounds, the nanoflow workflow reduced the biomass required for detection by approximately 20- to >600-fold, with pronounced gains for low-abundance metabolites such as NADPH and acetyl-CoA. TBA ion-pairing also enabled trap-and-elute nanoflow analysis of retained polar metabolites from single-cell-equivalent inputs. ATP was detected from one cell equivalent using both full-scan and targeted parallel reaction monitoring acquisition, with targeted acquisition further increasing signal over blank. Finally, we applied the workflow to stable isotope tracing in uniformly labeled 13C-glucose-treated cells. 13C-labeled ATP isotopologues were detectable from single-cell-equivalent input, and targeted acquisition improved isotopologue measurement near the detection limit. Together, these results demonstrate that nanoflow TBA ion-pairing LC-MS enables retained, high-sensitivity analysis of polar metabolites from ultra-low inputs and provides a foundation for extending central carbon metabolite analysis and isotope tracing toward single-cell-scale applications.

biochemistry↗

bioRxiv2026

PALINCODE: Recording cell lineage with ternary palindromic CRISPR bits

Reconstructing complete and accurate lineage trees remains a long-standing challenge in biology. Here, we introduce PALINCODE (Palindromic Coding and Decoding), a system that utilizes ternary CRISPR bits (cBits) to stochastically write one of three possible states over time, permanently embedding lineage relationships in the genome. We demonstrate PALINCODEs lineage-recording potential through simulations and establish palindromic CRISPR editing in cell culture models. We show that truncated Cas9 guide sequences yield ternary outcomes at high efficiency when compared to conventional guides. Using PALINCODE, we derived lineage-recording cell lines with a theoretical coding capacity of up to 10^25 bits, enabling the generation of lineage trees 32 cell divisions deep in single-cell sequencing of 293T cells. Furthermore, we applied PALINCODE using an in vivo melanoma model to jointly read out lineage history and gene expression, enabling in vivo reconstruction of clonal evolution within tumor cell clonal populations. PALINCODE circumvents several limitations of prior CRISPR-based systems while increasing the information potential at individual CRISPR sites, creating a lineage-recording platform with higher density than many competing approaches.

genomics↗

bioRxiv2026

Hierarchical Phase-Contrast Tomography Imaging: Applicability in biomedical research

ObjectivesHierarchical Phase-Contrast Tomography (HiP-CT) enables non-destructive, multi-scale imaging of whole human organs. We describe how HiP-CT is utilized for biomedical research within the Human Organ Atlas Hub through three case studies: mapping the enteric nervous system (ENS) of the human colon, analysing myocardial and AV conduction architecture in Tetralogy of Fallot (TOF), and characterizing ductal organization in breast carcinoma. The challenges we faced with this novel biomedical data are discussed. MethodsWhole-organ and region-of-interest scans of three types of human organs were acquired at the European Synchrotron Radiation Facility (ESRF) with isotropic voxel sizes ranging from 20 {micro}m to 0.8 {micro}m. For the colon, voxel binning and RootPainter were employed to tackle data size to segment the ENS. For the heart, voxel-wise myocyte orientation mapping was calculated in terabyte-scale datasets with a high-performance computational framework (Cardiotensor). Breast carcinoma samples were correlated with histopathology for structure validation. ResultsHiP-CT revealed the large-scale organization of the ENS in the colon, enabling visualisation of the 3D structures of the ENS across the colon In TOF hearts, analysis uncovered abnormal myocardial structure and heterogeneous conduction system morphology. In breast carcinoma, HiP-CT resolved the full hierarchy of ductal structures and vascular relationships within tumour and peritumoral regions. ConclusionsHiP-CT provides unprecedented, hierarchical insight into intact human organ structure, bridging the gap between histology and radiology. Advances in knowledgeHiP-CT establishes a new ex vivo radiological modality capable of linking microscale pathology to whole-organ context, advancing translational research in neurogastroenterology, cardiology, and oncology

biophysics↗

bioRxiv2026

A Pan-Cancer Single-Cell Compendium of Intratumoural Heterogeneity.

Intratumoural heterogeneity remains one of the most formidable challenges in oncology, driving treatment resistance, disease progression, and poor patient outcomes. To uncover the cellular programs that define tumour composition and diversity, we interrogated over 3.6 million single cells derived from more than 1,000 primary and metastatic tumours spanning 14 diverse cancer types. Through the identification and quantification of coordinated gene expression programs, we delineated heterogeneous cancer cell states, exhibiting unique and clinically significant associations with patient survival. Integration with high-resolution spatial transcriptomics identified prognostic cancer cell states marked by aggressive transcriptional programs, co-localizing with invasive histological features and significantly enriched in metastatic settings. Functional drug perturbation screening uncovered drug vulnerabilities specific to distinct cancer cell states, supporting both direct targeting strategies and therapeutic reprogramming toward less aggressive cellular phenotypes. Collectively, this study offers novel insights into the relationship between intratumoural transcriptional heterogeneity, spatial organization, clinical outcomes, and therapeutic vulnerabilities.

cancer biology↗

bioRxiv2025

Myosin VI orchestrates estrogen-driven gene expression in breast cancer cells.

Approximately 75% of breast cancer cases are estrogen receptor (ER)-positive, with endocrine therapies forming the foundation of treatment. However, therapeutic resistance remains a major clinical challenge, necessitating the identification of new molecular targets. Myosin VI (MVI), a motor protein increasingly linked to cancer, directly interacts with the ER and plays a key role in the spatial regulation of RNA Polymerase II. Notably, expression levels of MVI and ER are positively correlated across breast cancer tissues. Here, we present a multidisciplinary investigation combining advanced imaging, genomic profiling, and phenotypic characterisation to elucidate the interplay between MVI and the ER. We demonstrate that MVI nuclear localisation is dynamically regulated by estrogen signalling and ER activity. Conversely, ER nuclear localisation requires active MVI, suggesting a reciprocal regulatory mechanism. Furthermore, MVI influences subnuclear architecture, modulating ER transcriptional activity and downstream gene expression programmes that govern cell proliferation and migration. Importantly, pharmacological inhibition of MVI enhances the effect of hormone therapy, resulting in greater disruption of ER function than monotherapy alone. Moreover, MVI inhibition also suppresses the activity of hormone-resistant ER mutants, highlighting its potential to overcome therapy resistance. Our findings establish MVI as a critical regulator of ER nuclear dynamics and gene expression, supporting its candidacy as a novel therapeutic target in ER-positive breast cancer.

cancer biology↗

bioRxiv2025

Structural basis for constitutive activation of oncogenic gp130 mutants in human inflammatory hepatocellular adenomas

Inflammatory hepatocellular adenomas (IHCAs) are benign liver tumors primarily driven by somatic mutations in the gp130 receptor, resulting in its constitutive activation. To uncover the structural mechanisms underlying this activation, we employed cryo-electron microscopy and mutagenesis to study five representative IHCAs-associated gp130 mutants and two engineered mutants with ligand-independent activity. Most mutants formed "X"-shaped dimers through a 3D domain-swapping mechanism, characterized by large juxtamembrane separations and reliant on D1 domain-mediated "tip-to-tip" interactions for dimer clustering and activation. A rare mutant displayed a distinct "Y"-shaped dimer conformation, with closely aligned juxtamembrane domains enabling direct activation independent of dimer clustering. These findings highlight the diversity of gp130 mutant activation mechanisms and provide a foundation for developing therapeutic strategies targeting aberrant gp130 signaling in IHCAs.

biochemistry↗

bioRxiv2025

Ascending nociceptive pathways drive rapid escape and sustained avoidance in adult Drosophila

Nociception -- the detection of harmful stimuli by the nervous system -- contributes to both rapid escape and long-term avoidance behaviors. Drosophila larvae detect damaging heat, mechanical, and chemical stimuli with specialized multidendritic (md) neurons, and these cells are among the only sensory neurons that survive metamorphosis. However, it remains unknown which somatosensory neurons contribute to nociception in adult flies. In an optogenetic screen, we found that abdominal md neurons were the only somatosensory class to induce rapid escape and sustained place avoidance. Calcium imaging from abdominal md axons revealed that they are activated by thermal nociceptive stimuli (>40{degrees}C). Connectomic reconstruction showed that md axons form their strongest synaptic connections with ascending interneurons that project to the brain. Among these, we identified two classes of ascending neurons that mediate rapid escape responses and a third that supports sustained avoidance. Our findings reveal that adult Drosophila meet several core criteria commonly used to define pain: dedicated nociceptors, ascending pathways connecting peripheral sensors to integrative brain centers, and a capacity for sustained avoidance of noxious stimuli.

neuroscience↗

bioRxiv2025

Proprioceptive limit detectors mediate sensorimotor control of the Drosophila leg

Many animals possess mechanosensory neurons that fire when a limb nears the limit of its physical range, but the function of these proprioceptive limit detectors remains poorly understood. Here, we investigate a class of proprioceptors on the Drosophila leg called hair plates. Using calcium imaging in behaving flies, we find that a hair plate on the fly coxa (CxHP8) detects the limits of anterior leg movement. Reconstructing CxHP8 axons in the connectome, we found that they are wired to excite posterior leg movement and inhibit anterior leg movement. Consistent with this connectivity, optogenetic activation of CxHP8 neurons elicited posterior postural reflexes, while silencing altered the swing-to-stance transition during walking. Finally, we use comprehensive reconstruction of peripheral morphology and downstream connectivity to predict the function of other hair plates distributed across the fly leg. Our results suggest that each hair plate is specialized to control specific sensorimotor reflexes that are matched to the joint limit it detects. They also illustrate the feasibility of predicting sensorimotor reflexes from a connectome with identified proprioceptive inputs and motor outputs.

neuroscience↗

bioRxiv2025

Phylogeny-driven design of broadly protective sarbecovirus receptor-binding domain nanoparticle vaccines

Vaccines against emerging SARS-CoV-2 variants and sarbecoviruses with pandemic potential must elicit a robust humoral immune response in a population imprinted with the SARS-CoV-2 spike (S) protein. Here, we designed protein nanoparticle (NP) vaccines co-displaying the SARS-CoV-2 BA.5, SARS-CoV-1, and BtKY72 receptor-binding domains (RBDs) with or without the Wuhan-Hu-1 (Wu) RBD. We show that these vaccines elicit cross-reactive and broadly neutralizing plasma antibody responses against SARS-CoV-2 variants and sarbecoviruses in naive and pre-immune animals. Immunization with multivalent RBD-NPs overcomes immune imprinting and elicits neutralizing antibodies and memory B cells specific for the BA.5, SARS-CoV-1, and BtKY72 RBDs in mRNA-1273-vaccinated non-human primates. Multivalent RBD-NPs outperform a monovalent Wu RBD-NP vaccine by providing superior protection in mice and non-human primates challenged with the vaccine-mismatched SARS-CoV-2 XBB.1.5 or the pre-emergent RsSHC014. These data support the use of multivalent RBD-NP vaccines for SARS-CoV-2 variants and sarbecoviruses in naive and pre-immune populations.

immunology↗

bioRxiv2025

BASELINE: A CRISPR Base Editing Platform for Mammalian-Scale Single-Cell Lineage Tracing

A cells fate is shaped by its inherited state, or lineage, and the ever-shifting context of its environment. CRISPR-based recording technologies are a promising solution to map the lineage of a developing system, yet challenges remain regarding single-cell recovery, engineering complexity, and scale. Here, we introduce BASELINE, which uses base editing to generate high-resolution lineage trees in conjunction with single-cell profiling. BASELINE uses the Cas12a adenine base editor to irreversibly edit nucleotides within 50 synthetic target sites, which are integrated multiple times into a cells genome. We show that BASELINE accumulates lineage-specific marks over a wide range of biologically relevant intervals, recording more than 4300 bits of information in a model of pancreatic cancer, a 50X increase over existing technologies. Single-cell sequencing reveals high-fidelity capture of these recorders, recovering lineage reconstructions up to 40 cell divisions deep, within the estimated range of mammalian development. We expect BASELINE to apply to a wide range of lineage-tracing projects in development and disease, especially in which cellular engineering makes small, more distributed systems challenging.

synthetic biology↗

bioRxiv2024

Investigating the potential role of capsaicin in facilitating the spread of coxsackievirus B3 via extracellular vesicles

Coxsackievirus B3 (CVB3) is a non-enveloped picornavirus that can cause systemic inflammatory diseases including myocarditis, pericarditis, pancreatitis, and meningoencephalitis. We have previously reported that following infection, CVB3 localizes to mitochondria, inducing mitochondrial fission and mitophagy, while inhibiting lysosomal degradation by blocking autophagosome-lysosome fusion. This results in the release of virus-laden mitophagosomes from the host cell as infectious extracellular vesicles (EVs) which allow non-lytic viral egress. Transient receptor potential vanilloid 1 (TRPV1/TRPV1) is a heat and capsaicin-sensitive cation channel that regulates mitochondrial dynamics by inducing mitochondrial membrane depolarization and fission. In this study, we found that treating cells with the TRPV1 agonist capsaicin dramatically enhances CVB3 egress via EVs. Analysis of the released EVs revealed increased levels of viral capsid protein VP1/VP1, mitochondrial protein TOM70/TOMM70, and fission protein phospho-DRP1/DNM1L (Ser 616). Moreover, these EVs exhibited increased levels of heat shock protein HSP70/HSPA1A, suggesting a potential role of these chaperones in facilitating infectious EV release from cells. Furthermore, TRPV1 inhibition with capsazepine significantly reduced viral infection in vitro. We previously observed similar effects in vitro with another TRPV1 inhibitor SB-366791. Our current in vivo studies found that SB-366791 significantly mitigates pancreatic damage and reduces viral titers in mouse model of CVB3 pancreatitis. Given the lack of understanding regarding the factors that contribute to diverse clinical manifestations of CVB3, our study highlights capsaicin and TRPV1 as potential exacerbating factors that facilitates CVB3 dissemination via mitophagy-derived EVs. IMPORTANCECVB3 is a prevalent pathogen responsible for a range of severe diseases, including myocarditis, pericarditis, pancreatitis, and meningoencephalitis. Despite its clinical significance, factors that determine the severity of CVB3 infection and why some individuals experience life-threatening manifestations while others have mild, cold-like symptoms remain poorly understood. This study provides new insights into the molecular mechanisms underlying CVB3 dissemination and pathogenesis. By investigating the role of capsaicin, a common dietary component, in modulating viral spread, we demonstrate that activation of TRPV1 by capsaicin enhances release of infectious CVB3 via mitophagy-derived EVs. Our results offer novel evidence that modulating TRPV1 activity could influence the clinical outcomes of CVB3 infection, opening new avenues for therapeutic interventions. Given the widespread consumption of capsaicin, this study highlights an important dietary factor that could play a role in shaping CVB3 pathogenesis and its clinical manifestations, underscoring the potential for targeted strategies to mitigate severe disease outcomes.

microbiology↗

bioRxiv2024

Anti-HIV-1 B cell antigen receptor signaling and structure

The B cell antigen receptor (BCR) complex, comprised of antigen recognition and signaling components, functions in initiating B cell activation. While structural studies have described BCR domain organization, gaps remain in our understanding of its antigen binding domain (Fab, fragment antigen-binding) disposition, and how antigen binding is sensed to initiate signaling. Here, we report antigen affinity and signaling of the immunoglobulin (Ig) class IgM and IgG BCRs and define conformational states of full-length BCRs of two human broadly neutralizing antibodies, the glycan-specific, heavy chain domain-swapped, I-shaped 2G12, and a canonical Y-shaped antibody, CH31, that recognizes the CD4-binding site on the HIV-1 Envelope protein (Env). The BCRs adopted the shapes (I or Y) of their respective soluble antibodies, and both Ig class of BCRs of the same specificity bound Env trimers with similar affinities. We observed antigen-valenc y dependent differential signaling by the 2G12 IgM and IgG BCRs with trimeric Envs. Cryo-electron microscopy of the 2G12 IgG and CH31 IgM BCRs revealed varied Fab orientations. Structural comparisons revealed hinge points and regions of flexibility in the BCRs suggesting a highly dynamic structure of the BCR complex. Taken together, our results provide an integrated understanding of BCR structure, conformation, antigen recognition and signaling, and provide the basis for understanding antigen induced BCR signal transmission. One Sentence SummaryCryo-electron microscopy structures of full-length B cell antigen receptor complex provide a novel dynamic BCR model and a basis for understanding antigen binding induced signal transmission.

immunology↗

bioRxiv2024

Mechano-osmotic signals control chromatin state and fate transitions in pluripotent stem cells

Acquisition of specific cell shapes and morphologies is a central component of cell fate transitions. Although signaling circuits and gene regulatory networks that regulate pluripotent stem cell differentiation have been intensely studied, how these networks are integrated in space and time with morphological transitions and mechanical deformations to control state transitions remains a fundamental open question. Here, we focus on two distinct models of pluripotency, primed pluripotent stem cells and pre-implantation inner cell mass cells of human embryos to discover that cell fate transitions associate with rapid changes in nuclear shape and volume which collectively alter the nuclear mechanophenotype. Mechanistic studies in human induced pluripotent stem cells further reveal that these phenotypical changes and the associated active fluctuations of the nuclear envelope arise from growth factor signaling-controlled changes in chromatin mechanics and cytoskeletal confinement. These collective mechano-osmotic changes trigger global transcriptional repression and a condensation-prone environment that primes chromatin for a cell fate transition by attenuating repression of differentiation genes. However, while this mechano-osmotic chromatin priming has the potential to accelerate fate transitions and differentiation, sustained biochemical signals are required for robust induction of specific lineages. Our findings uncover a critical mechanochemical feedback mechanism that integrates nuclear mechanics, shape and volume with biochemical signaling and chromatin state to control cell fate transition dynamics.

cell biology↗

bioRxiv2024

Chemigenetic far-red labels and Ca2+ indicators optimized for photoacoustic imaging.

Photoacoustic imaging is an emerging modality with significant promise for biomedical applications such as neuroimaging, owing to its capability to capture large fields of view, deep inside complex scattering tissue. However, the widespread adoption of this technique has been hindered by a lack of suitable molecular reporters for this modality. In this work, we introduce chemigenetic labels and calcium sensors specifically tailored for photoacoustic imaging, using a combination of synthetic dyes and HaloTag-based self-labelling proteins. We rationally design and engineer far-red "acoustogenic" dyes, showing high photoacoustic turn-ons upon binding to HaloTag, and develop a suite of tunable calcium indicators based on these scaffolds. These first-generation photoacoustic reporters show excellent performance in tissue-mimicking phantoms, with the best variants outperforming existing sensors in terms of signal intensity, sensitivity and photostability. We demonstrate the application of these ligands for labelling HaloTag-expressing neurons in mouse brain tissue, producing strong, specifically targeted photoacoustic signal, and provide a first example of in vivo labelling with these chemigenetic photoacoustic probes. Together, this work establishes a new approach for the design of photoacoustic reporters, paving the way towards deep tissue functional imaging.

biochemistry↗

bioRxiv2024

A single-cell atlas characterizes dysregulation of the bone marrow immune microenvironment associated with outcomes in multiple myeloma

Multiple Myeloma (MM) remains incurable despite advances in treatment options. Although tumor subtypes and specific DNA abnormalities are linked to worse prognosis, the impact of immune dysfunction on disease emergence and/or treatment sensitivity remains unclear. We established a harmonized consortium to generate an Immune Atlas of MM aimed at informing disease etiology, risk stratification, and potential therapeutic strategies. We generated a transcriptome profile of 1,149,344 single cells from the bone marrow of 263 newly diagnosed patients enrolled in the CoMMpass study and characterized immune and hematopoietic cell populations. Associating cell abundances and gene expression with disease progression revealed the presence of a proinflammatory immune senescence-associated secretory phenotype in rapidly progressing patients. Furthermore, signaling analyses suggested active intercellular communication involving APRIL-BCMA, potentially promoting tumor growth and survival. Finally, we demonstrate that integrating immune cell levels with genetic information can significantly improve patient stratification.

genomics↗

bioRxiv2024

Tuning a mechanical model to biological reality: A case study in the LaMSA system of the trap-jaw ant Strumigenys

Understanding the relationship between morphology and movement in biomechanical systems, particularly those composed of multiple complex elements, presents challenges due to the nonlinear nature of the interaction between components. This study focuses on the mandibular closing mechanisms in ants, specifically comparing muscle-driven actuation (MDA) and latch-mediated spring actuation (LaMSA) in the genus Strumigenys. Analyzing 3D structural data from diverse Strumigenys species, we employ mathematical models for both LaMSA and MDA systems. Our findings reveal distinct patterns of mechanical sensitivity between the two models, with sensitivity varying across kinematic output metrics. We explore the performance transition between MDA and LaMSA systems by incorporating biological data and correlations between morphological parameters into the models. In these models tuned specifically to Strumigenys, we find the LaMSA mechanism outperforms MDA at small relative mandible mass. Notably, the location and abruptness of the performance transition differs among various kinematic performance metrics. Overall, this work contributes a novel approach to understanding form-function relationships in complex biomechanical systems. By using morphological data to calibrate a general biomechanical model for a particular group, it strikes a balance between simplicity and specificity and allows for conclusions that are uniquely tuned to the morphological characteristics of the group.

biophysics↗