Biology subjects

Hempel, S.

Publications and source records attributed to Hempel, S..

9 recordsLinked to original sources

bioRxiv2026

Immunomodulatory impact of PTPN11/SHP2-based vertical RAS-MAPK pathway inhibition in pancreatic cancer

Protein tyrosine phosphatase non-receptor 11 (PTPN11/SHP2) is a critical upstream mediator of RAS-MAPK signaling and a central node in adaptive resistance mechanisms evolving with RAS and MEK/ERK inhibition. Accordingly, clinical trials are currently evaluating allosteric SHP2 inhibitors in vertical RAS pathway combination therapies for various KRAS-mutant malignancies, including pancreatic ductal adenocarcinoma (PDAC). Here, we aimed to delineate the immunomodulatory effects of SHP2-based vertical RAS pathway inhibition in notoriously immunotherapy-refractory PDAC, spanning from early treatment response to invariably evolving adaptive resistance. Employing human and murine PTPN11 knockout and wild-type PDAC cell lines, an autochthonous murine PDAC model (KPC), and patient-derived PDAC organoids, we find that short term dual MEK/SHP2 inhibition induces increased T cell infiltration and a reduction in immunosuppressive M2-like macrophages. However, these effects are accompanied by a decrease in mature dendritic cells and a concomitant expansion of monocytic myeloid-derived suppressor cells, indicative of a mixed immunological response with both immune-activating and immune-suppressing features. These changes are associated with tumor cell-intrinsic upregulation of CXCR3 ligands and TGF-{beta}, as well as increased expression of checkpoint ligands for TIGIT and TIM-3 across molecular subtypes and species. With prolonged treatment and transition to an adaptive resistant tumor cell state, the initial immune-sensitizing effects are lost and the immune-suppressive features prevail. M2-like macrophages re-accumulate, dendritic cell maturation remains impaired, TGF-{beta} expression persists, and TIGIT and TIM-3 ligand expression is further enhanced. Notably, dual SHP2/RAS inhibition recapitulates the observed induction of TGF-{beta} and checkpoint ligands. Collectively, these findings identify a dynamic but ultimately immunosuppressive remodeling of the tumor microenvironment in response to SHP2-based vertical RAS pathway inhibition in PDAC and provide a rationale for combinatorial immunotherapy strategies. In particular, concurrent targeting of TGF-{beta}, combined TIGIT/TIM-3 checkpoint blockade, and likely CD40 agonism may help sustain early immune activation while counteracting emerging suppressive features, thereby improving the durability of tumor control.

immunology↗

bioRxiv2025

What are all these soil fungi doing? Complex carbon use ability as a predictive trait for fungal community functions

Fungal communities in soil play important roles in decomposition processes and soil organic carbon cycling. These communities are tremendously diverse, making it challenging to assign relevant functions to individual species. Fungal communities may be differentiated at the level of functional guilds; beyond such broad classification we have little delimitation, especially in fungal taxa common in grassland and agricultural soils. To resolve the level of functional similarity in fungal communities and define traits predictive of soil carbon cycling, we characterized fungal isolates abundant in agricultural soils to test the hypotheses that (i) the majority of saprobic soil fungi have the ability to use complex carbon sources, (ii) differences in complex carbon use abilities correlate with fungal enzymatic profiles, following principles of the fungal economics spectrum and (iii) carbon use ability is a predictive trait for fungal community functions. Using specialized growth media, we isolated and characterized 105 isolates and developed a novel FungiResp approach that directly tests fungal activity on complex carbon sources. The largest amount of variance between isolates was explained by differential abilities to use cellulose and starch, with only few phylogenetically distinct fungal clades showing high respiratory activity on these biopolymers. A preference for bacterial necromass was another major distinction among taxa. These key traits correlated with soil fungal community shifts in response to carbon substrate availability. By contrast, enzymatic activity was a poor predictor of fungal carbon use ability, except for correlations in lignin use and laccase activity. The newly established functional trait of carbon use ability offers important insights into diverse fungal communities: Many taxa lack the ability to use complex carbon (on their own), while the most common enzymes analyzed in soil showed little correlation with fungal mineralization potential. The discovery of key functional traits is an important step towards predicting the significance of fungal community shifts for soil carbon cycling.

ecology↗

bioRxiv2024

Revisiting soil fungal biomarkers and conversion factors: Interspecific variability in phospholipid fatty acids, ergosterol and rDNA copy numbers

The abundances of fungi and bacteria in soil are used as simple predictors for carbon dynamics, and represent widely available microbial traits. Soil biomarkers serve as quantitative estimates of these microbial groups, though not quantifying microbial biomass per se. The accurate conversion to microbial carbon pools, and an understanding of its comparability among soils is therefore needed. We refined conversion factors for classical fungal biomarkers, and evaluated the application of quantitative PCR (qPCR, rDNA copies) as a biomarker for soil fungi. Based on biomarker contents in pure fungal cultures of 30 isolates, combined with available references, we propose average conversion factors of 95.3 g fungal C g-1 ergosterol, 32.0 mg fungal C {micro}mol-1 PLFA 18:2{omega}6,9 and 0.264 pg fungal C ITS1 DNA copy-1. As expected, interspecific variability was most pronounced in rDNA copies, though qPCR results showed the least phylogenetic bias. A modeling approach based on exemplary agricultural soils further supported the hypothesis that high diversity in soil buffers against biomarker variability, whereas also phylogenetic biases impact the accuracy of comparisons in biomarker estimates. Our analyses suggest that qPCR results cover the fungal community in soil best, though with a variability only partly offset in highly diverse soils. PLFA 18:2{omega}6,9 and ergosterol represent accurate biomarkers to quantify Ascomycota and Basidiomycota. To conclude, the ecological interpretation and coverage of biomarker data prior to their application in global models is important, where the combination of different biomarkers may be most insightful.

ecology↗

bioRxiv2023

Deep learning image analysis for filamentous fungi taxonomic classification: Dealing with small data sets with class imbalance and hierarchical grouping

Deep learning applications in taxonomic classification for animals and plants from images have become popular, while those for microorganisms are still lagging behind. Our study investigated the potential of deep learning for the taxonomic classification of hundreds of filamentous fungi from colony images, which is typically a task that requires specialized knowledge. We isolated soil fungi, annotated their taxonomy using standard molecular barcode techniques, and took images of the fungal colonies grown in petri dishes (n = 606). We applied a convolutional neural network with multiple training approaches and model architectures to deal with some common issues in ecological datasets: small amounts of data, class imbalance, and hierarchically structured grouping. Model performance was overall low due mainly to the relatively small data set, class imbalance, and the high morphological plasticity exhibited by fungal colonies. However, our approach indicates that morphological features such as color, patchiness, and colony extension rate, could be used for the recognition of fungal colonies at higher taxonomic ranks (i.e., phylum, class, and order). Model explanation implies that image recognition characters appear at different positions within the colony (e.g., outer or inner hyphae), depending on the taxonomic resolution. Our study suggests the potential of deep learning applications for better understanding the taxonomy and ecology of filamentous fungi amenable to axenic culturing. Meanwhile, our study also highlights some technical challenges in deep learning image analysis in ecology, highlighting that the domain of applicability of these methods needs to be carefully considered.

microbiology↗

bioRxiv2021

Non-mycorrhizal root associated fungi of a tropical montane forest are relatively robust to the long-term addition of moderate rates of nitrogen and phosphorus

Andean forests are biodiversity hotspots and globally important carbon (C) repositories. This status might be at risk due to increasing rates of atmospheric nutrient deposition. As fungal communities are key in the recirculation of soil nutrients, assessing their responses to soil eutrophication can help establish a link between microbial biodiversity and the sustainability of the C sink status of this region. Beyond mycorrhizal fungi, which have been studied more frequently, a wide range of other fungi associate with the fine root fraction of trees. Monitoring these communities can offer insights into how communities composed of both facultative and obligate root associated fungi are responding to soil eutrophication. Here we document the response of non-mycorrhizal root associated fungal (RAF) communities to a long-term nutrient manipulation experiment. The stand level fine root fraction of an old growth tropical montane forest was sampled after seven years of nitrogen (N) and phosphorus (P) additions. RAF communities were characterized by a deep sequencing approach. As per the resource imbalance model, we expected that asymmetries in the availability of C, N and P elicited by fertilization will lead to mean richness reductions and alterations of the community structure. We recovered moderately diverse fungal assemblages composed by sequence variants classified within a wide set of trophic guilds. While mean richness remained stable, community composition shifted, particularly among Ascomycota and after the addition of P. Fertilization factors, however, only accounted for a minor proportion of the variance in community composition. These findings suggest that, unlike mycorrhizal fungi, RAF communities are less sensitive to shifts in soil nutrient availability. A plausible explanation is that non-mycorrhizal RAF have fundamentally different nutrient acquisition and life history traits, thus allowing them greater stoichiometric plasticity and an array of functional acclimation responses that collectively express as subtle shifts in community level attributes.

microbiology↗

bioRxiv2021

Disentangling the relative importance of spatio-temporal parameters and host selectivity in shaping AMF communities in temperate forests

Many woody and herbaceous plants in temperate forests cannot establish and survive in the absence of mycorrhizal associations. Here, we address and hierarchize how Glomeromycota, a group of soil-borne fungi, forming a ubiquitous type of mycorrhiza, varied in a temperate forest patch in Germany with time, space, plant hosts but also the proximity to Glomeromycota-associating woody species. The communities of Glomeromycota in our study were non-random. We observed that space had a greater impact on fungal community structure than either time or proximity to Glomeromycota-associating trees but unlike host identity did not alter Glomeromycotan richness. The set of parameters which we addressed has rarely been studied together and we believe that the resulting ranking could ease prioritizing some of them to include in future surveys. Glomeromycota are crucial for the establishment of understory plants in temperate forests making it desirable to further explore how they vary in time and space.

ecology↗

bioRxiv2018

Evidence for subsoil specialization in arbuscular mycorrhizal fungi

Arbuscular mycorrhizal (AM) fungal communities are now known to vary with depth in arable land. Here we use two previously published high-throughput Illumina sequencing data sets, and compare a 52 year long chronosequence of recultivated agriculture fields after a topsoil and subsoil mixing event, with a set of undisturbed topsoil and subsoil samples from a similar field. We show that AM taxa identified as subsoil indicators are exclusively present in early stages of the chronosequence, whereas topsoil indicator taxa can be found across the chronosequence, and that similarities from the chronosequence fields to the subsoil communities decrease with time. Our results provide evidence on the ecological specialization of certain AM fungal taxa to deep soil layers.

ecology↗

bioRxiv2017

Mir-132 controls beta cell proliferation and survival in mouse model through the PTEN/AKT/FOXO3 signaling

Aim and hypothesismicroRNAs (miRNAs) play an integral role in maintaining beta cell function and identity. Deciphering their targets and precise role, however, remains a challenge. In this study we aimed to identify miRNAs and their downstream targets involved in regeneration of islet beta cells following partial pancreatectomy in mice.\n\nMethodsRNA from laser capture microdissected (LCM) islets of partially pancreatectomized and sham-operated mice were profiled with microarrays to identify putative miRNAs implicated in control of beta cell regeneration. Altered expression of selected miRNAs, including miR-132, was verified by RT-PCR. Potential targets of miR-132 were seleced through bioinformatic data mining. Predicted miR-132 targets were validated for their changed RNA and protein expression levels and signaling upon miR-132 knockdown or overexpression in MIN6 cells. The ability of miR-132 to foster beta cell proliferation in vivo was further assessed in pancreatectomized miR-132-/- and control mice.\n\nResultsPartial pancreatectomy significantly increased the number of BrdU+/insulin+ positive islet cells. Microarray profiling revealed 14 miRNAs, including miR-132 and -141, to be significantly upregulated in LCM islets of partially pancreatectomized compared to LCM islets of control mice. In the same comparison miR-760 was the only miRNA found to be downregulated. Changed expression of these miRNAs in islets of partially pancreatectomized mice was confirmed by RT-PCR only in the case of miR-132 and -141. Based on previous knowledge of its function, we chose to focus our attention on miR-132. Downregulation of miR-132 in MIN6 cells reduced proliferation while enhancing the expression of proapoptic genes, which was instead reduced in miR-132 overexpression MIN6 cells. Microarray profiling, RT-PCR and immunoblotting of miR-132 overexpressing MIN6 cells revealed their downregulated expression of Pten, with concomitant increased levels of pro-proliferative factors phospho-Akt and phospho-Creb as well as inactivation of pro-apoptotic Foxo3 via its phosphorylation. Finally, we show that regeneration of beta cells following partial pancreatectomy was reduced in miR-132-/- mice compared to control mice.\n\nConclusions/InterpretationsOur study provides compelling evidence for upregulation of miR-132 being critical for regeneration of mouse islet beta cells in vivo through downregulation of its target Pten. Hence, the miR-132/Pten/Akt/Foxo3 signaling pathway may represent a suitable target to enhance beta cell mass.\n\nResearch in ContextWhat is already known?\n\nO_LISeveral miRNAs, including miR-132, are known to regulate beta cell function and mass in several mouse models of diabetes db/db, ob/ob and high fat-diet.\nC_LI\n\nWhat is the key question?\n\nO_LIWhich are miRNAs implicated in control of beta cell regeneration upon partial pancreatectomy and how?\nC_LI\n\nWhat are the new findings?\n\nO_LImiR-132 is critical to promote regeneration of mouse beta cells in vivo following partial pancreatectomy\nC_LIO_LIIn vitro studies in mouse MIN6 cells indicate that miR-132 fosters beta cell proliferation by down-regulating the expression of phosphatase Pten, thereby tilting the balance between anti-apoptotic factor Akt and pro-apoptotic factor Foxo3 activities towards proliferation through regulation of their phosphorylation.\nC_LI\n\nHow might this impact on clinical practice in the foreseeable future?\n\nO_LIThese findings strengthen the rationale for targeting the expression of miR-132 to increase beta cell mass in vivo (type 2 diabetes) or ex-vivo (islet transplantation in type 1 diabetes) for the treatment of diabetes.\nC_LI

cell biology↗

bioRxiv2017

Subsoil arbuscular mycorrhizal fungal communities in arable soil differ from those in topsoil

Arbuscular mycorrhizal fungi are recognized as important drivers of plant health and productivity in agriculture but very often existing knowledge is limited to the topsoil. With growing interest in the role of subsoil in sustainable agriculture, we used high-throughput Illumina sequencing on a set of samples encompassing drilosphere, rhizosphere and bulk soil, in both top- and subsoil. Our results show subsoil AMF communities harbor unique Operational Taxonomic Units (OTUs) and that both soil depths differ in community structure both at the OTU and family level. Our results emphasize the distinctness of subsoil AMF communities and the potential role of subsoil as a biodiversity reservoir.

ecology↗