Cachexia represents a fatal energy-wasting syndrome in a large number of patients with cancer that mostly results in a pathological loss of skeletal muscle and adipose tissue. Here we show that tumor ...cell exposure and tumor growth in mice triggered a futile energy-wasting cycle in cultured white adipocytes and white adipose tissue (WAT), respectively. Although uncoupling protein 1 (Ucp1)-dependent thermogenesis was dispensable for tumor-induced body wasting, WAT from cachectic mice and tumor-cell-supernatant-treated adipocytes were consistently characterized by the simultaneous induction of both lipolytic and lipogenic pathways. Paradoxically, this was accompanied by an inactivated AMP-activated protein kinase (Ampk), which is normally activated in peripheral tissues during states of low cellular energy. Ampk inactivation correlated with its degradation and with upregulation of the Ampk-interacting protein Cidea. Therefore, we developed an Ampk-stabilizing peptide, ACIP, which was able to ameliorate WAT wasting in vitro and in vivo by shielding the Cidea-targeted interaction surface on Ampk. Thus, our data establish the Ucp1-independent remodeling of adipocyte lipid homeostasis as a key event in tumor-induced WAT wasting, and we propose the ACIP-dependent preservation of Ampk integrity in the WAT as a concept in future therapies for cachexia.
Abstract
Definiens and Mosaic Laboratories developed an immuno-oncology (IO) panel consisting of multiplex chromogenic IHC assays and image and data analysis solutions to determine a standardized ...profile of the tumor microenvironment in non-small cell lung cancer (NSCLC). Expression of PD-L1 on tumor and immune cells, the density of infiltrating immune cell populations, and their spatial pattern have been shown to be predictive for response to immunotherapy. These factors are currently being evaluated to determine whether they can guide treatment across different categories of immunotherapies (e.g., checkpoint inhibition, vaccination, CAR-T cell therapy, or combinations thereof). Multiplex chromogenic IHC assays were developed for PDL1/CD68/CD3 and FoxP3/PD-1/CD8 and single-stain IHC was used to detect cells that were positive for granzyme B. Image analysis were optimized to detect biomarker positive cells (e.g., PD-L1+, CD3-, CD68-), double positive cells (e.g., PD-L1+, CD3-, CD68+), negative cells, or percent of area positive for Granzyme B. Densities for each cell population were quantified and pathologist annotations were used to distinguish between tumor core and invasive margin regions. Machine learning with random forests and decision trees was used to automatically distinguish tumor and stroma compartments to evaluate differences in densities of infiltrating immune cells. Densities of the different cell populations and their spatial relationships were analyzed to assign cases to one of the following immune status categories: acquired immune resistance, immune tolerance, adaptive checkpoint inhibitor resistance (all three are subtypes of high immune cell density), impaired infiltration, immune ignorance, intrinsic checkpoint inhibitor resistance (subtypes of low immune cell density). NSCLC tissues were analyzed and examples of biomarker profiles for each category are shown. The distribution of immune categories between squamous and non-squamous NSCLC subtypes and a comparison of immune cell densities and spatial relationships are shown. We conclude that the IO panel can be used to establish standardized immune profiles in NCCLC. In the future, it can be used to search for novel prognostic and predictive signatures and to establish the prevalence of patients with defined biomarker profiles in different subsets of patients (e.g, different stages, morphologic subtypes, or mutational status).
Citation Format: Lisa M. Dauffenbach, Christopher A. Kerfoot, Gela Sia, Anthony Masci, Johannes Zimmermann, Jan Lesniak, Alexei Budco, Svenja Lippok, Katrin Schneider, René Korn, Tobias Wiestler, Dasa Medrikova, Florian Leiß. Characterization of inflammatory cell patterns and densities using multiplex immunohistochemistry immuno-oncology assays abstract. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2017 Oct 26-30; Philadelphia, PA. Philadelphia (PA): AACR; Mol Cancer Ther 2018;17(1 Suppl):Abstract nr B069.
Abstract Background: Immune checkpoint inhibitors (ICIs) targeting PD-(L)1 following concurrent chemoradiotherapy (cCRT) have shown clinical activity in locally advanced, unresectable NSCLC, e.g. ...with the PACIFIC regimen 1. PD-L1 expression is typically assessed by pathologist scoring of immunohistochemically (IHC) stained tissue using the tumor cell (TC) score. In this work, we present PD-L1 Quantitative Continuous Scoring (PD-L1 QCS) to digitally assess PD-L1 expression (Ventana SP263 assay), and analyze its utility for identification of patient subgroups with higher likelihood of treatment response. Method: 368 digitized whole slide images (WSI) of fresh biopsy and archival samples were obtained from the PACIFIC trial (NCT02125461), including 126 patients who received placebo and 242 patients who received Durvalumab post-cCRT 1. WSI analysis included two deep learning models for epithelial TC segmentation followed by optical density (OD) quantification of membrane expression 2. Initially, PD-L1 QCS was calibrated to compute the TC score at 1% cutoff by determining an optimal OD cell positivity threshold. Next, other scoring approaches were investigated, which included various OD thresholds and biomarker cutoffs. Results: PD-L1 QCS-based TC scoring was able to maintain median overall survival (mOS) of 57.4 months in the biomarker positive (BM+) ICI cohort, while increasing prevalence to 191 out of 242 patients (78.9%) compared to 177 (73.1%) with manual TC score at 1% cutoff. Extending the scoring paradigm to the PD-L1+ TC density >= 542.5 cells/mm² lead to a more pronounced increase of prevalence to 200 patients (82.6%) in the ICI population, while mOS was not reached and additionally showed comparable hazard ratios (HR) when evaluating against the BM+ placebo cohort (HR=0.54 0.39, 0.76 resp. HR=0.52 0.37, 0.74). Results further demonstrated that PD-L1 QCS density scoring of the ICI treated population allowed for a clearer separation of patients with less treatment benefit (34.2m mOS, N=42, log rank p=0.0067) compared to manual scoring (47.4m mOS, N=65, log rank p=0.2). Conclusion: We investigated a computational pathology approach for selecting patients for treatment with the PACIFIC regimen and compared it against established manual pathology scoring. Results indicate that PD-L1 QCS TC proportion scoring can increase prevalence, while PD-L1 QCS TC density scoring has the potential to identify an even larger patient subgroup with increased survival benefit. References: 1. Antonia, Scott J., et al. "Durvalumab after chemoradiotherapy in stage III non-small-cell lung cancer.” New England Journal of Medicine 377.20 (2017): 1919-1929. 2. Lesniak, Jan, et al. " Quantitative computational assessment of PD-L1 enables robust patient selection for biomarker-informed anti-PD-L1 treatment of NSCLC patients." J. Immunother. Cancer, Vol. 10., 2022. Citation Format: Jan Martin Lesniak, Ina Bisha, Simon Christ, Lina Meinecke, Thomas Padel, Laura Sebastian Monasor, Pallavi Sontakke, Tze Heng Tan, Dasa Medrikova, Andrea Storti, Anatoliy Shumilov, Hadassah Sade, Günter Schmidt, Ross Stewart, Yashaswi Shrestha. Quantitative continuous scoring of PD-L1 for superior patient selection for anti-PD-L1 treatment of locally advanced or unresectable NSCLC abstract. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7617.
The n‐3 polyunsaturated fatty acids, especially eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), exert hypolipidemic effects and prevent development of obesity and insulin resistance in ...animals fed high‐fat diets. We sought to determine the efficacy of α‐substituted DHA derivatives as lipid‐lowering, antiobesity, and antidiabetic agents. C57BL/6 mice were given a corn oil‐based high‐fat (35% weight/weight) diet (cHF), or cHF with 1.5% of lipids replaced with α‐methyl DHA ethyl ester (Substance 1), α‐ethyl DHA ethyl ester (Substance 2), α,α‐di‐methyl DHA ethyl ester (Substance 3), or α‐thioethyl DHA ethyl ester (Substance 4) for 4 months. Plasma markers of glucose and lipid metabolism, glucose tolerance, morphology, tissue lipid content, and gene regulation were characterized. The cHF induced obesity, hyperlipidemia, impairment of glucose homeostasis, and adipose tissue inflammation. Except for Substance 3, all other substances prevented weight gain and Substance 2 exerted the strongest effect (63% of cHF‐controls). Glucose intolerance was significantly prevented (∼67% of cHF) by both Substance 1 and Substance 2. Moreover, Substance 2 lowered fasting glycemia, plasma insulin, triacylglycerols, and nonesterified fatty acids (73, 9, 47, and 81% of cHF‐controls, respectively). Substance 2 reduced accumulation of lipids in liver and skeletal muscle, as well as adipose tissue inflammation associated with obesity. Substance 2 also induced weight loss in dietary obese mice. In contrast to DHA administered either alone or as a component of the EPA/DHA concentrate (replacing 15% of dietary lipids), Substance 2 also reversed established glucose intolerance in obese mice. Thus, Substance 2 represents a novel compound with a promising potential in the treatment of obesity and associated metabolic disturbances.
Departments of 1 Adipose Tissue Biology, 2 Bioenergetics, and 3 Neurophysiology of Memory and Computational Neuroscience, Institute of Physiology, Academy of Sciences of the Czech Republic; and 4 ...Department of Physiology and Developmental Biology, Faculty of Science, Charles University, Prague, Czech Republic
Submitted 27 February 2008
; accepted in final form 20 May 2008
The obesogenic effect of a high-fat (HF) diet is counterbalanced by stimulation of energy expenditure and lipid oxidation in response to a meal. The aim of this study was to reveal whether muscle nonshivering thermogenesis could be stimulated by a HF diet, especially in obesity-resistant A/J compared with obesity-prone C57BL/6J (B/6J) mice. Experiments were performed on male mice born and maintained at 30°C. Four-week-old mice were randomly weaned onto a low-fat (LF) or HF diet for 2 wk. In the A/J LF mice, cold exposure (4°C) resulted in hypothermia, whereas the A/J HF, B/6J LF, and B/6J HF mice were cold tolerant. Cold sensitivity of the A/J LF mice was associated with a relatively low whole body energy expenditure under resting conditions, which was normalized by the HF diet. In both strains, the HF diet induced uncoupling protein-1-mediated thermogenesis, with a stronger induction in A/J mice. Only in A/J mice: 1 ) the HF diet augmented activation of whole body lipid oxidation by cold; and 2 ) at 30°C, oxygen consumption, total content, and phosphorylation of AMP-activated protein kinase (AMPK), and AICAR-stimulated palmitate oxidation in soleus muscle was increased by the HF diet in parallel with significantly increased leptinemia. Gene expression data in soleus muscle of the A/J HF mice indicated a shift from carbohydrate to fatty acid oxidation. Our results suggest a role for muscle nonshivering thermogenesis and lipid oxidation in the obesity-resistant phenotype of A/J mice and indicate that a HF diet could induce thermogenesis in oxidative muscle, possibly via the leptin-AMPK axis.
nonshivering thermogenesis; leptin; adenosine monophosphate-activated protein kinase
Address for reprint requests and other correspondence: J. Kopecky, Dept. of Adipose Tissue Biology, Inst. of Physiology, Academy of Sciences of the Czech Republic, Videnska 1083, 142 20 Prague, Czech Republic (e-mail: kopecky{at}biomed.cas.cz )
Abstract
Many targeted cancer therapies rely on biomarkers, which are assessed by standard pathologist scoring of immunohistochemically stained tissue. However, this process is subjective, ...semi-quantitative and does not assess expression heterogeneity. A quantitative method to measure IHC markers might therefore significantly improve patient selection particularly of proteins expressed at low levels. To address these challenges, we have developed the Quantitative Continuous Scoring (QCS) that deploys the power of fully supervised Deep Learning (DL) algorithms to provide objective and continuous data of biomarkers in digitized IHC whole slide images (WSI). The two DL-based algorithms, developed using pathologist input as the ground truth, identify areas of invasive tumor and segment each individual tumor cell across the WSI into pixels that represent cell nuclei, cytoplasm and/or membrane. This allows to compute biomarker expression as mean Optical Density (OD) in each of these subcellular compartments based on the Hue-Saturation-Density (HSD) model. Of note, this also allows computation of the spatial distribution of tumor cells across the WSI. The measured OD for each cell is aggregated as a histogram to quantitative continuous readouts for each patient sample. The method’s ability to accurately detect low expression range facilitates selection of antibody clones for IHC assays, has been successfully used to delineate mode of action and PK/PD mechanisms, has provided surrogate markers of spatial expression heterogeneity to predict potential bystander activity and has facilitated marker co-expression analysis to inform rational combination therapies. In retrospective clinical trials analysis, QCS showed superior performance in identifying a patient population gaining maximum treatment benefit. QCS-based quantification of PD-L1 membrane expression was able to stratify anti-PD-L1 treated late-stage non-small cell lung cancer (NSCLC) patients NCT01693562 with a higher prevalence and more significant log rank p-value (64%, p=0.0001) for OS compared to pathologist TPS (59%, p=0.01). In summary, we describe a computational pathology-based approach for precise biomarker quantification and superior patient selection with broad applicability and the potential to transform pathology, thus addressing one of the key challenges of precision oncology.
Citation Format: Hadassah Sade, Ansh Kapil, Philipp Wortmann, Andreas Spitzmueller, Nicolas Triltsch, Lina Meinecke, Susanne Haneder, Anatoliy Shumilov, Jan Lesniak, Valeria Bertani, Tze-Heng Tan, Ana Hidalgo-Sastre, Simon Christ, Andrea Storti, Regina Alleze, Dasa Medrikova, Jessica Chan, Simon Lanzmich, Markus Schick, Guenter Schmidt, J. Carl Barrett. Quantitative assessment of IHC using computational pathology allows superior patient selection for biomarker-informed patients abstract. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 468.
OBJECTIVE--The induction of obesity, dyslipidemia, and insulin resistance by high-fat diet in rodents can be prevented by n-3 long-chain polyunsaturated fatty acids (LC-PUFAs). We tested a hypothesis ...whether AMP-activated protein kinase (AMPK) has a role in the beneficial effects of n-3 LC-PUFAs. RESEARCH DESIGN AND METHODS--Mice with a whole-body deletion of the alpha2 catalytic subunit of AMPK (AMPKalpha.sup.2.sup.-/- ) and their wild-type littermates were fed on either a low-fat chow, or a corn oil-based high-fat diet (cHF), or a cHF diet with 15% lipids replaced by n-3 LC-PUFA concentrate (cHF+F). RESULTS--Feeding a cHF diet induced obesity, dyslipidemia, hepatic steatosis, and whole-body insulin resistance in mice of both genotypes. Although cHF+F feeding increased hepatic AMPKalpha2 activity, the body weight gain, dyslipidemia, and the accumulation of hepatic triglycerides were prevented by the cHF+F diet to a similar degree in both AMPKalpha2.sup.-/- and wildtype mice in ad libitum-fed state. However, preservation of hepatic insulin sensitivity by n-3 LC-PUFAs required functional AMPKalpha2 and correlated with the induction of adiponectin and reduction in liver diacylglycerol content. Under hyperinsulinemic-euglycemic conditions, AMPKalpha2 was essential for preserving low levels of both hepatic and plasma triglycerides, as well as plasma free fatty acids, in response to the n-3 LC-PUFA treatment. CONCLUSIONS--Our results show that n-3 LC-PUFAs prevent hepatic insulin resistance in an AMPKalpha2-dependent manner and support the role of adiponectin and hepatic diacylglycerols in the regulation of insulin sensitivity. AMPKalpha2 is also essential for hypolipidemic and antisteatotic effects of n-3 LC-PUFA under insulin-stimulated conditions. Diabetes 59:2737-2746, 2010
Regulatory T (Treg) cells are critical determinants of both immune responses and metabolic control. Here we show that systemic ablation of Treg cells compromised the adaptation of whole-body energy ...expenditure to cold exposure, correlating with impairment in thermogenic marker gene expression and massive invasion of pro-inflammatory macrophages in brown adipose tissue (BAT). Indeed, BAT harbored a unique sub-set of Treg cells characterized by a unique gene signature. As these Treg cells respond to BAT activation upon cold exposure, this study defines a BAT-specific Treg sub-set with direct implications for the regulation of energy homeostasis in response to environmental stress.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Mitochondrial uncoupling protein 2 (UCP2) is abundant in developing monocyte/macrophage cells and may affect hematopoiesis by reducing formation of reactive oxygen species. The aims of this study ...were to further characterize the involvement of UCP2 in hematopoiesis.
In situ hybridization in mouse embryos identified UCP2-positive cells in liver and inside primitive blood vessels from 10.5 days of prenatal development. High UCP2 transcript levels were detected in reticulocytes and other maturating erythroid cells in peripheral blood of mice exposed to hypoxia, and in umbilical cord blood of human neonates and peripheral blood of adults. Our results suggest involvement of UCP2 in erythropoiesis.
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