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Nobis, M.

Publications and source records attributed to Nobis, M..

10 recordsLinked to original sources

Environmental drivers of heritable trait variation and lag of adaptation to climate in Hordeum murinum

1. Most plant species are genetically differentiated among populations, often reflected by phenotypic trait variation that corresponds to local adaptation. Yet the strength of local adaptation and heritable contribution to phenotypic traits vary across traits, species, and environments. Additionally, climate change is rapidly altering environmental conditions, and the climate may shift faster than populations can adapt or track the change via dispersal, resulting in adaptive lags. However, it remains unclear how widespread such adaptive lags are across plant species. 2. We focused on Hordeum murinum, an annual ruderal grass widespread in Europe. We combined continental-scale in situ measurements of 2070 plants across 207 populations with common garden experiments across two contrasting climates and two soil types to disentangle heritable variation from phenotypic plasticity and assess potential adaptive lags under climate change. 3. We found that heritable variation was pronounced in developmental traits, particularly flowering time and plant height, while seed weight, reproductive investment and SLA showed intermediate heritable contribution, and flag leaf area and total biomass were primarily plastic. Heritable trait variation was strongly associated with temperature at the populations origin, and trait clines were consistent with in situ patterns, suggesting that temperature is the main driver of genetic differentiation in H. murinum. However, we detected that fitness peaked in populations originating from warmer climates, indicating that evolutionary responses may not keep pace with rapid environmental shifts. 4. Synthesis: Our results highlight that H. murinum harbors substantial heritable variation, shaped primarily by temperature. However, the pace of evolutionary change may be insufficient to track ongoing climate change, leaving populations potentially vulnerable to future environmental conditions.

ecology↗

Pulsed priming with the FAK inhibitor narmafotinib enhances both gemcitabine/Abraxane and FOLFIRINOX chemotherapy response in pancreatic cancer

BackgroundPancreatic ductal adenocarcinoma (PDAC) is a particularly lethal malignancy with few treatment options available. Extensive remodelling of extracellular matrix (ECM) generates a highly fibrotic tumour landscape, which impairs therapeutic response. ObjectiveWe investigated whether stromal priming via the highly specific Focal Adhesion Kinase (FAK) inhibitor narmafotinib (AMP945) in combination with the two major standard-of-care chemotherapies in PDAC, gemcitabine/Abraxane and FOLFIRINOX, reduces fibrosis and enhances treatment efficacy. Design3D organotypic matrices, intravital imaging, and in vivo subcutaneous and orthotopic PDAC models were used to provide a rationale for a first-line priming regimen of narmafotinib prior to chemotherapy. ResultsNeoadjuvant chemotherapy induces fibrosis in PDAC indicating a need for upfront first-line priming of the ECM to normalise the stroma for optimal treatment response. Narmafotinib is a new potent small molecule FAK inhibitor. Phase I safety data shows excellent safety, tolerability, and pharmacokinetics following oral administration in humans. We reveal that narmafotinib treatment during early ECM remodelling ( priming) reduces fibrosis, while limiting subsequent PDAC invasion. Moreover, intravital imaging demonstrates real-time FAK inactivation and cell cycle stalling, leading to improved chemotherapeutic efficacy upon narmafotinib priming in vivo. Long-term assessment in patient-derived models shows that narmafotinib priming prior to gemcitabine/Abraxane or FOLFIRINOX reduces PDAC progression and extends survival in both chemotherapy settings. ConclusionsOur results using these Phase II-ready drug combinations strongly support the clinical assessment of narmafotinib in PDAC. Narmafotinib is currently in Phase Ib/IIa trials, assessing a pulsed dosing regimen prior to gemcitabine/Abraxane, and warrants further clinical assessment in combination with FOLFIRINOX. SIGNIFICANCE OF THIS STUDYO_ST_ABSWhat is already known on this topicC_ST_ABSO_LIPancreatic cancer (PC) is one of the most lethal malignancies and is characterised by a dense, fibrotic stroma, which impairs chemotherapy efficacy. C_LIO_LIThe non-receptor tyrosine kinase FAK is known to promote cancer fibrosis and therefore represents a therapeutic target to normalise the PC stroma and to improve chemotherapy performance. C_LI What this study addsO_LINeoadjuvant chemotherapy induces early fibrosis indicating a need for upfront first-line priming of the ECM to blunt or normalise stromal fibrosis for optimal response to therapy. C_LIO_LIThe small molecule inhibitor narmafotinib (which is currently under Phase Ib/IIa clinical trial assessment) shows high specificity towards FAK as well as desirable pharmacokinetics and pharmacodynamics in healthy human volunteers. C_LIO_LIEarly short-term narmafotinib priming reduces fibrosis and improves the efficacy of subsequent standard-of-care gemcitabine/Abraxane chemotherapy. C_LIO_LIFOLFIRINOX (oxaliplatin, irinotecan, leucovorin and 5-fluorouracil) is a multi-agent chemotherapy preferentially used in PDAC patients with good performance status. Our results demonstrate that narmafotinib priming also improves FOLFIRINOX efficacy, leading to extended survival in patient-derived PDAC models. C_LI How this study might affect research, practice, or policyO_LIThis study supports the clinical development of narmafotinib in combination with both gemcitabine/Abraxane (ACCENT trial) and further FOLFIRINOX standard-of-care chemotherapies for PDAC patient treatment. C_LIO_LIThe first-line priming strategy and early ECM normalisation used in this study may also be applicable to other combination therapy settings and warrants further investigation in ongoing clinical studies. C_LI

cancer biology↗

Climate-driven in-situ trait variation in an annual ruderal grass across Europe

O_LIPlant functional traits link environmental conditions to plant performance and adaptation. Growing evidence shows that in-situ intraspecific trait variation can be as important as differences between species, yet large intraspecific datasets measured in situ are rare. While most studies focused on plant morphological traits, contents of elemental nutrients in the seeds received much less attention so far. C_LIO_LIWe conducted a large-scale in-situ study of the widespread ruderal grass Hordeum murinum. We sampled 2070 individuals from 207 populations from a large part of the native range in Europe and Northern Africa. In-situ, we measured seed ripening phenology and morphological traits, and analyzed the content of elemental nutrients in the seeds. C_LIO_LIWe found that Hordeum murinum grew larger, produced seeds later, and had heavier seeds in colder and wetter areas. Plants in denser vegetation were taller with heavier seeds but formed fewer spikes. Seed nutrient content generally declined with seed weight and was mainly driven by climate. Soil conditions had only minor effects on plant traits and seed nutrients. Population identity explained much of the variation, indicating a possible genetic component. C_LIO_LISynthesis: Our findings provide a comprehensive view of Hordeum murinum responding to environmental gradients across its European distribution. Climatic variables, especially temperature, are key drivers for reproductive success and seed nutrient content, while local environments, such as biotic pressures, are more critical for growth-related traits. These patterns indicate that H. murinum modulates its growth and reproductive investment along environmental gradients, balancing phenology, stress tolerance, and limited competitive capacity. C_LI

ecology↗

ROCK2 inhibition has a dual role in reducing ECM remodelling and cell growth, while impairing migration and invasion

The Rho-associated coiled-coil containing protein kinases 1/2 (ROCK1/2) are key signalling proteins involved in the regulation of the actin cytoskeleton and control a variety of cellular processes. This includes cell proliferation, stemness, cell migration and invasion as well as actomyosin contraction and extracellular matrix (ECM) remodelling. Due to a lack of ROCK2 specific inhibitors, previous pharmacological studies of ROCK function have relied on pan-ROCK1/2 inhibition. Here, we use the ROCK2 specific inhibitor, GV101, in the context of breast cancer (BC) models to uncouple the effects of ROCK2 pharmacological inhibition on both epithelial and stromal fibroblast cell populations. We demonstrate that ROCK2 inhibition with GV101 reduces fibroblast-mediated remodelling of pre-existing and de novo synthesised ECM resulting in a reduction in biomechanical stiffness, while also limiting epithelial cell growth in 2D and 3D settings. We also demonstrate that ROCK2 inhibition exposes epithelial cell vulnerability to fluid flow-induced shear stress. Furthermore, using 3D co-cultures we reveal that GV101 reduces single-cell migratory capacity, while the disruption of stromal ECM architecture is sufficient to impede collective cell migration and invasion. Finally, we assessed ROCK2 expression in human BC patient tumours revealing that ROCK2 expression is up-regulated during BC progression and that high ROCK2 expression correlates with poor patient outcome in the triple-negative subtype of BC. Together, these results demonstrate that ROCK2 specific inhibition disrupts key functions in both epithelial cell and stromal fibroblast compartments warranting further assessment of ROCK2 in highly proliferative, fibrotic or metastatic diseases, such as BC. HighlightsO_LIFibroblast-mediated remodelling of pre-existing and de novo synthesised extracellular matrix is reduced upon ROCK2 specific targeting. C_LIO_LIROCK2 inhibition impairs epithelial cancer cell growth and colony formation in 2D and 3D contexts, while also limiting single-cell migration and cell survival upon fluid flow-induced shear stress. C_LIO_LIROCK2 targeting during fibroblast-mediated ECM remodelling is sufficient to disrupt collective cell migration as well as 3D invasion into organotypic fibroblast-contracted collagen matrices. C_LIO_LIROCK2 expression is up-regulated during breast cancer progression and high expression correlates with poor patient outcome in triple-negative breast cancer. C_LI

cell biology↗

A selective inhibitor of oncogenic JNK signalling perturbs metastatic outgrowth of triple-negative breast cancer through metabolic blockade

Although c-Jun N-terminal Kinase (JNK) represents an attractive anti-cancer target, its pleiotropic functionality limits the use of direct JNK inhibitors. Here, we identify a distinct subcellular pattern of JNK activity as a therapeutic vulnerability in breast cancer, where cytoplasmic JNK activity predicts poor survival outcomes, is elevated in triple-negative breast cancers (TNBC) and is essential for metastatic outgrowth. Mechanistic analyses reveal cytoplasmic JNK acts through multiple mechanisms, with downstream targets involved in cellular metabolism and cytoskeletal regulation. On this basis, we leveraged actin-based phenotypic drug-screening and identified K12, an indirect but selective inhibitor of cytoplasmic JNK that blocks TNBC metastatic outgrowth in vivo. We reveal that K12 inhibits glutaminase-1 and the pyruvate dehydrogenase complex, and that this poly-pharmacology overcomes pyruvate anaplerosis, a known resistance mechanism of existing glutaminase inhibitors. These findings demonstrate the potential of selectively targeting the oncogenic function of JNK, offering new treatment options for early-stage metastatic TNBC.

cancer biology↗

Live microscopy of multicellular spheroids with the multi-modal near-infrared nanoparticles reveals differences in oxygenation gradients

Assessment of hypoxia, nutrients, metabolite gradients, and other hallmarks of the tumor microenvironment within 3D multicellular spheroid and organoid models represents a challenging analytical task. Here, we report red/near-infrared emitting cell staining O2-sensitive nanoparticles, which enable measurements of spheroid oxygenation on a conventional fluorescence microscope. Nanosensor probes, termed MMIR (multi-modal infrared), incorporate a near-infrared O2-sensitive metalloporphyrin (PtTPTBPF) and a deep red aza-BODIPY reference dyes within a biocompatible polymer shell, allowing oxygen gradients quantification via fluorescence ratio and phosphorescence lifetime readouts. We optimized staining techniques and evaluated nanosensor probe characteristics and cytotoxicity. Subsequently, we applied nanosensors to the live spheroid models based on HCT116, DPSCs, and SKOV3 cells, at rest and treated with drugs affecting cell respiration. We found that the growth medium viscosity, spheroids size, and formation method influenced spheroid oxygenation. Unexpectedly, some spheroids (produced from HCT116 and dental pulp stem cells) exhibited inverted oxygenation gradients, with higher core oxygen levels than the periphery. This contrasted with the frequently encountered normal gradient of hypoxia towards the core caused by diffusion. Further microscopy analysis of spheroids with an "inverted" gradient demonstrated metabolic stratification of cells within spheroids: thus, autofluorescence FLIM of NAD(P)H revealed the formation of glycolytic core, and localization of OxPhos-active cells at the periphery. Collectively, we demonstrate a strong potential of NIR-emitting ratiometric nanosensors for advanced microscopy studies targeting live and quantitative real-time monitoring of cell metabolism and hypoxia in complex 3D tissue models. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=38 SRC="FIGDIR/small/571110v2_ufig1.gif" ALT="Figure 1"> View larger version (14K): org.highwire.dtl.DTLVardef@49cef8org.highwire.dtl.DTLVardef@1b31072org.highwire.dtl.DTLVardef@fcaf4aorg.highwire.dtl.DTLVardef@1583b5f_HPS_FORMAT_FIGEXP M_FIG C_FIG

cancer biology↗

An autophagy program that promotes T cell egress from the lymph node controls responses to immune checkpoint blockade

Lymphatic endothelial cells (LECs) lining the lymphatic vessels of the lymph node (LN) parenchyma orchestrate leukocyte trafficking and peripheral T cell dynamics. T cell responses to immunotherapy largely rely on peripheral T cell recruitment in tumors. Yet, a systematic and molecular understanding of how LECs within the LNs control T cell dynamics under steady state and tumor-bearing conditions is lacking. Using intravital and high-resolution imaging combined with immune phenotyping, we show that LEC-specific deletion of the essential autophagy gene Atg5 alters intranodal positioning of lymphocytes and accrues their persistence in the LNs, by increasing the availability of the main egress signal S1P. Single-cell RNA-sequencing of tumor-draining LNs from WT and ATG5LEC-KO mice unveils that loss of ATG5 remodels niche-specific LEC phenotypes, involved in molecular pathways regulating lymphocyte trafficking and LEC-T cell interactions. Functionally, loss of LEC-autophagy prevents recruitment of tumor-infiltrating T cells and NK cells and abrogates tumor regression in response to anti-PD-1 or anti-CTLA4-based immunotherapy. Thus, a unique LEC-autophagy program boosts immune-checkpoint responses by guiding systemic T cell dynamics. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=143 SRC="FIGDIR/small/549282v1_ufig1.gif" ALT="Figure 1"> View larger version (51K): org.highwire.dtl.DTLVardef@1a4bfe1org.highwire.dtl.DTLVardef@1317fe2org.highwire.dtl.DTLVardef@c2fa20org.highwire.dtl.DTLVardef@b35dba_HPS_FORMAT_FIGEXP M_FIG C_FIG

immunology↗

A Microtubule Mechanostat Enables Cells to Navigate Confined Environments

Cells migrating through complex 3D environments experience considerable physical challenges including tensile stress and compression. To move, cells need to resist these forces whilst also squeezing the large nucleus through confined spaces. This requires highly coordinated cortical contractility. Microtubules can both resist compressive forces and sequester key actomyosin regulators to ensure appropriate activation of contractile forces. Yet, how these two roles are integrated to achieve nuclear transmigration in 3D is largely unknown. Here, we demonstrate that compression triggers reinforcement of a dedicated microtubule structure at the rear of the nucleus by the mechanoresponsive recruitment of CLASPs (cytoplasmic linker-associated proteins) which dynamically strengthens and repairs the lattice. These reinforced microtubules form the mechanostat: an adaptive feedback mechanism that allows the cell to both withstand compressive force and spatiotemporally organise contractility signalling pathways. The microtubule mechanostat facilitates nuclear positioning and coordinates force production to enable the cell to pass through constrictions. Disruption of the mechanostat imbalances cortical contractility, stalling migration and ultimately resulting in catastrophic cell rupture. Our findings reveal a new role for microtubules as cellular sensors which detect and respond to compressive forces, enabling movement and ensuring survival in mechanically demanding environments. One Sentence SummaryMechanically tuned microtubules form a mechanostat to coordinate contractility and nuclear positioning in confined migration.

cell biology↗

Resistance to platinum chemotherapy in lung adenocarcinoma is driven by a non-genetic, cell-cycle dependent mechanism

We previously used a pulse-based in vitro assay to unveil targetable signalling pathways associated with innate cisplatin resistance in lung adenocarcinoma (Hastings et al., 2020). Here we advanced this model system and identified a non- genetic mechanism of resistance that drives recovery and regrowth in a subset of cells. Using RNAseq and a suite of biosensors to track single cell fates both in vitro and in vivo, we identified that early S phase cells have a greater ability to maintain proliferative capacity, which correlated with reduced DNA damage over multiple generations. In contrast, cells in G1, late S or those treated with PARP/RAD51 inhibitors, maintained higher levels of DNA damage and underwent prolonged S/G2 phase arrest and senescence. Combined with our previous work, these data indicate that there is a non-genetic mechanism of resistance in lung adenocarcinoma that is dependent on the cell cycle stage at the time of cisplatin exposure.

cell biology↗

Molecular phylogenetics and micromorphology of Australasian Stipeae (Poaceae), and the interrelation of whole-genome duplication and evolutionary radiations in this grass tribe

The mainly Australian grass genus Austrostipa with ca. 64 species represents a remarkable example of an evolutionary radiation. To investigate aspects of diversification, macro- and micromorphological variation in this genus we conducted a molecular phylogenetic and scanning electron microscopy (SEM) analysis including representatives from all of its accepted subgenera. Plastid DNA variation within Austrostipa was low and only few lineages were resolved. Nuclear ITS and Acc1 yielded comparable groupings of taxa and resolved subgenera Arbuscula, Petaurista, Bambusina in a common clade and as monophyletic. In summary, we suggest recognizing nine subgenera in Austrostipa. Because of its taxonomic significance in Stipeae, we studied the lemma epidermal structure in 34 representatives of Austrostipa. In most species, the lemma epidermal pattern (LEP) was relatively uniform (maize-like LEP), but in six species it was more similar to that of Stipa s.str., Neotrinia, Ptilagrostis and Orthoraphium. The species representing subgenera Lobatae, Petaurista, Bambusina and A. muelleri from subg. Tuberculatae were well-separated from all the other species included in the analysis. Two different sequence copies of Acc1 were found in polyploid Austrostipa and Anemanthele. Each of the copy types formed a single clade. This was also true of the sampled species of Stipa s.str., but their clades were strongly separated from those of Austrostipa and Anemanthele. This underlines the statement of Tzvelev (1977) that most if not all contemporary Stipeae are of hybrid origin and demonstrates it for the first time unambiguously on the molecular level. Chromosome number variation is surveyed and reviewed for the whole tribe Stipeae and interpreted in a molecular phylogenetic framework. The rather coherent picture of chromosome number variation underlines the phylogenetic and evolutionary significance of this character. The closest extant relatives of Austrostipa and Anemanthele are in the clade of Achnatherum s.str., Celtica, Oloptum and Stipellula. These genera are most abundant in Central and Eastern Asia, which makes a colonization of Australian and New Zealand from this region more likely, perhaps via long-distance dispersal, than colonization of Australia from southern South America via Antarctica as previously invoked. Supporting Information may be found online in the Supporting Information section at the end of the article.

plant biology↗