bims-cagime 2023-07-16 papers

bims-cagime

Biomed News

on Cancer, aging and metabolism

Issue of 2023–07–16
33 papers selected by
Kıvanç Görgülü, Technical University of Munich

Ethanol inhibits pancreatic acinar cell autophagy through upregulation of ATG4B, mediating pathologic responses of alcoholic pancreatitis.
A Ketogenic Diet in Combination with Gemcitabine Mitigates Pancreatic Cancer-Associated Cachexia in Male and Female KPC Mice.
Dual Recombinase-Based Mouse Models Help Decipher Cancer Biology and Targets for Therapy.
The Troyer syndrome protein spartin mediates selective autophagy of lipid droplets.
A mouse model for high-efficient Flp-recombinase-mediated genetic manipulation in the pancreas.
Prolonged fasting times reap greater geroprotective effects when combined with caloric restriction in adult female mice.
ATPase activity of DFCP1 controls selective autophagy.
Raman Imaging Reveals Insights into Membrane Phase Biophysics in Cells.
You complete me: tumor cell-myeloid cell nuclear fusion as a facilitator of organ-specific metastasis.
Spectrum of Response to Platinum and PARP Inhibitors in Germline BRCA-Associated Pancreatic Cancer in the Clinical and Preclinical Setting.
Fluorescence microscopy imaging of mitochondrial metabolism in cancer cells.
Chemically induced reprogramming to reverse cellular aging.
Ribonucleotide synthesis by NME6 fuels mitochondrial gene expression.
Pancreatic ductal adenocarcinoma induces neural injury that promotes a transcriptomic and functional repair signature by peripheral neuroglia.
How circulating tumor cluster biology contributes to the metastatic cascade: from invasion to dissemination and dormancy.
Transcriptional Profile of Human Pancreatic Acinar Ductal Metaplasia.
Corylifol A ameliorates muscle atrophy by inhibiting TAOK1/p38-MAPK/FoxO3 pathway in cancer cachexia.
OME-Zarr: a cloud-optimized bioimaging file format with international community support.
Protocol for evaluation of tumor-derived exosome-induced cancer cell metastasis in a mouse model.
Small extracellular vesicles promote stiffness-mediated metastasis.
Tethering ATG16L1 or LC3 induces targeted autophagic degradation of protein aggregates and mitochondria.
A Novel HOXA10-Associated 5 Gene-Based Prognostic Signature for Stratification of Short-Term Survivors of Pancreatic Ductal Adenocarcinoma.
Engineering RNA export for measurement and manipulation of living cells.
Genome-wide CRISPR screens define determinants of epithelial-mesenchymal transition mediated immune evasion by pancreatic cancer cells.
Physiological role of cytokines in the regulation of mammalian metabolism.
Modifiable Pancreatic Ductal Adenocarcinoma (PDAC) Risk Factors.
Oxidative phosphorylation is a metabolic vulnerability of endocrine therapy and palbociclib resistant metastatic breast cancers.
Real-Life Results of Palliative Chemotherapy in Metastatic Pancreatic Ductal Adenocarcinoma.
The stress sensor GCN2 differentially controls ribosome biogenesis in colon cancer according to the nutritional context.
A distinct stimulatory cDC1 subpopulation amplifies CD8+ T cell responses in tumors for protective anti-cancer immunity.
Evaluation and management of body composition changes in cancer patients.
Predictive value of baseline serum carbohydrate antigen 19-9 level on treatment effect of neoadjuvant chemoradiotherapy in patients with resectable and borderline resectable pancreatic cancer in two randomized trials.
Whole-body cellular mapping in mouse using standard IgG antibodies.

Am J Physiol Gastrointest Liver Physiol. 2023 Jul 11.

Ethanol inhibits pancreatic acinar cell autophagy through upregulation of ATG4B, mediating pathologic responses of alcoholic pancreatitis.

Olga A Mareninova, Sophie R Gretler, Grace E Lee, Michael Pimienta, Yueqiu Qin, Jason M Elperin, Jinliang Ni, Zsolt Razga, Anna S Gukovskaya, Ilya Gukovsky.

  Excessive alcohol intake is a major risk factor for pancreatitis, sensitizing exocrine pancreas to stressors by mechanisms that remain obscure. Impaired autophagy drives non-alcoholic pancreatitis, but the effects of ethanol (EtOH) and alcoholic pancreatitis on autophagy are poorly understood. Here, we find that ethanol reduces autophagosome formation in pancreatic acinar cells, both in a mouse model of alcoholic pancreatitis induced by combination of EtOH diet and cerulein (a CCK ortholog) and in EtOH+CCK treated acinar cells (ex-vivo model). Ethanol treatments decreased pancreatic level of LC3-II, a key mediator of autophagosome formation. This was caused by ethanol-induced upregulation of ATG4B, a cysteine protease which, cell-dependently, regulates the balance between cytosolic LC3-I and membrane-bound LC3-II. We show that ATG4B negatively regulates LC3-II in acinar cells subjected to EtOH treatments. Ethanol raised ATG4B level by inhibiting its degradation, enhanced ATG4B enzymatic activity, and strengthened its interaction with LC3-II. We also found ATG4B increase and impaired autophagy in a dissimilar, non-secretagogue model of alcoholic pancreatitis induced by EtOH plus palmitoleic acid. Adenoviral ATG4B overexpression in acinar cells greatly reduced LC3-II and inhibited autophagy. Further, it aggravated trypsinogen activation and necrosis, mimicking key responses of ex-vivo alcoholic pancreatitis. Conversely, shRNA Atg4B knockdown enhanced autophagosome formation and alleviated ethanol-induced acinar cell damage. The results reveal a novel mechanism whereby ethanol inhibits autophagosome formation and thus sensitizes to pancreatitis, and a key role of ATG4B in ethanol's effects on autophagy. Enhancing pancreatic autophagy, particularly by down-regulating ATG4B, could be beneficial in mitigating the severity of alcoholic pancreatitis.

Keywords:  CCK; acinar cells; alcohol; autophagy; pancreas

DOI:  https://doi.org/10.1152/ajpgi.00053.2023
Int J Mol Sci. 2023 Jun 28. pii: 10753. [Epub ahead of print]24(13):

A Ketogenic Diet in Combination with Gemcitabine Mitigates Pancreatic Cancer-Associated Cachexia in Male and Female KPC Mice.

Natalia E Cortez, Suraj Pathak, Cecilia Rodriguez Lanzi, Brian V Hong, Ryman Crone, Rasheed Sule, Fangyi Wang, Shuai Chen, Aldrin V Gomes, Keith Baar, Gerardo G Mackenzie.

  Cancer-associated cachexia (CAC) is a critical contributor to pancreatic ductal adenocarcinoma (PDAC) mortality. Thus, there is an urgent need for new strategies to mitigate PDAC-associated cachexia; and the exploration of dietary interventions is a critical component. We previously observed that a ketogenic diet (KD) combined with gemcitabine enhances overall survival in the autochthonous LSL-KrasG12D/+; LSL-Trp53 R172H/+; Pdx1-Cre (KPC) mouse model. In this study, we investigated the effect and cellular mechanisms of a KD in combination with gemcitabine on the maintenance of skeletal muscle mass in KPC mice. For this purpose, male and female pancreatic tumor-bearing KPC mice were allocated to a control diet (CD), a KD, a CD + gemcitabine (CG), or a KD + gemcitabine (KG) group. We observed that a KD or a KG-mitigated muscle strength declined over time and presented higher gastrocnemius weights compared CD-fed mice. Mechanistically, we observed sex-dependent effects of KG treatment, including the inhibition of autophagy, and increased phosphorylation levels of eIF2α in KG-treated KPC mice when compared to CG-treated mice. Our data suggest that a KG results in preservation of skeletal muscle mass. Additional research is warranted to explore whether this diet-treatment combination can be clinically effective in combating CAC in PDAC patients.

Keywords:  cachexia; cancer-associated cachexia; gemcitabine; ketogenic diet; pancreatic cancer

DOI:  https://doi.org/10.3390/ijms241310753
Cancer Res. 2023 Jul 14. 83(14): 2279-2282

Dual Recombinase-Based Mouse Models Help Decipher Cancer Biology and Targets for Therapy.

Tina Sket, Chiara Falcomatà, Dieter Saur.

The advent of next-generation sequencing (NGS) and single-cell profiling technologies has revealed the complex and heterogenous ecosystem of human tumors under steady-state and therapeutic perturbation. Breakthroughs in the development of genetically engineered mouse models (GEMM) of human cancers that are based on the combination of two site-specific recombinase systems [dual-recombinase system (DRS)] offer fundamental new possibilities to elucidate and understand critical drivers of the diverse tumor phenotypes and validate potential targets for therapy. Here, we discuss opportunities DRS-based cancer GEMMs offer to model, trace, manipulate, and functionally investigate established cancers, their interactions with the host, and their response to therapy.

DOI: https://doi.org/10.1158/0008-5472.CAN-22-2119
Nat Cell Biol. 2023 Jul 13.

The Troyer syndrome protein spartin mediates selective autophagy of lipid droplets.

Jeeyun Chung, Joongkyu Park, Zon Weng Lai, Talley J Lambert, Ruth C Richards, Jiuchun Zhang, Tobias C Walther, Robert V Farese.

Lipid droplets (LDs) are crucial organelles for energy storage and lipid homeostasis. Autophagy of LDs is an important pathway for their catabolism, but the molecular mechanisms mediating LD degradation by selective autophagy (lipophagy) are unknown. Here we identify spartin as a receptor localizing to LDs and interacting with core autophagy machinery, and we show that spartin is required to deliver LDs to lysosomes for triglyceride mobilization. Mutations in SPART (encoding spartin) lead to Troyer syndrome, a form of complex hereditary spastic paraplegia1. Interfering with spartin function in cultured human neurons or murine brain neurons leads to LD and triglyceride accumulation. Our identification of spartin as a lipophagy receptor, thus, suggests that impaired LD turnover contributes to Troyer syndrome development.

DOI: https://doi.org/10.1038/s41556-023-01178-w
Pancreatology. 2023 Jun 28. pii: S1424-3903(23)00190-4. [Epub ahead of print]

A mouse model for high-efficient Flp-recombinase-mediated genetic manipulation in the pancreas.

Xiaohui Zhu, Jiaxiang Chen, Bin Wang, Lingxiang Wang, Jiale Wang, Dongfeng Feng, Yan Yang, Oliver Wang, Ashley N Haddock, Ying Wang, Baoan Ji, Yan Bi.

   BACKGROUND: Tissue and cell-specific gene targeting has been widely employed in biomedical research. In the pancreas, the commonly used Cre recombinase recognizes and recombines loxP sites. However, to selectively target different genes in distinct cells, a dual recombinase system is required.
METHOD: We developed an alternative recombination system mediated by FLPo, which recognizes frt DNA sequences for pancreatic dual recombinase-mediated genetic manipulation. An IRES-FLPo cassette was targeted between the translation stop code and 3-UTR of the mouse pdx1 gene in a Bacterial Artificial Chromosome using recombineering technology. Transgenic BAC-Pdx1-FLPo mice were developed by pronuclear injection.
RESULTS: Highly efficient recombination activity was observed in the pancreas by crossing the founder mice with Flp reporter mice. When the BAC-Pdx1-FLPo mice were bred with conditional FSF-KRasG12D and p53 F/F mice, pancreatic cancer developed in the compound mice. The characteristics of pancreatic cancer resembled those derived from conditional LSL-KRasG12D and p53 L/L mice controlled by pdx1-Cre.
CONCLUSIONS: We have generated a new transgenic mouse line expressing FLPo, which enables highly efficient pancreatic-specific gene recombination. When combined with other available Cre lines, this system can be utilized to target different genes in distinct cells for pancreatic research.

Keywords:  Animal models; Cancer; Dual recombinase; Pancreas; Pdx1-FlpO

DOI:  https://doi.org/10.1016/j.pan.2023.06.013
Cell Metab. 2023 Jul 11. pii: S1550-4131(23)00180-8. [Epub ahead of print]35(7): 1179-1194.e5

Prolonged fasting times reap greater geroprotective effects when combined with caloric restriction in adult female mice.

Eleonora Duregon, Maria Emilia Fernandez, Jorge Martinez Romero, Clara Di Germanio, Meaghan Cabassa, Romaniya Voloshchuk, Margaux R Ehrlich-Mora, Jacqueline M Moats, Sarah Wong, Oye Bosompra, Annamaria Rudderow, Christopher H Morrell, Simonetta Camandola, Nathan L Price, Miguel A Aon, Michel Bernier, Rafael de Cabo.

  Emerging new evidence highlights the importance of prolonged daily fasting periods for the health and survival benefits of calorie restriction (CR) and time-restricted feeding (TRF) in male mice; however, little is known about the impact of these feeding regimens in females. We placed 14-month-old female mice on five different dietary regimens, either CR or TRF with different feeding windows, and determined the effects of these regimens on physiological responses, progression of neoplasms and inflammatory diseases, serum metabolite levels, and lifespan. Compared with TRF feeding, CR elicited a robust systemic response, as it relates to energetics and healthspan metrics, a unique serum metabolomics signature in overnight fasted animals, and was associated with an increase in lifespan. These results indicate that daytime (rest-phase) feeding with prolonged fasting periods initiated late in life confer greater benefits when combined with imposed lower energy intake.

Keywords:  aging phenotypes; calorie restriction; circadian misalignment; fasting; female mice; histopathology; metabolomics; time-restricted feeding

DOI:  https://doi.org/10.1016/j.cmet.2023.05.003
Nat Commun. 2023 Jul 08. 14(1): 4051

ATPase activity of DFCP1 controls selective autophagy.

Viola Nähse, Camilla Raiborg, Kia Wee Tan, Sissel Mørk, Maria Lyngaas Torgersen, Eva Maria Wenzel, Mireia Nager, Veijo T Salo, Terje Johansen, Elina Ikonen, Kay Oliver Schink, Harald Stenmark.

Cellular homeostasis is governed by removal of damaged organelles and protein aggregates by selective autophagy mediated by cargo adaptors such as p62/SQSTM1. Autophagosomes can assemble in specialized cup-shaped regions of the endoplasmic reticulum (ER) known as omegasomes, which are characterized by the presence of the ER protein DFCP1/ZFYVE1. The function of DFCP1 is unknown, as are the mechanisms of omegasome formation and constriction. Here, we demonstrate that DFCP1 is an ATPase that is activated by membrane binding and dimerizes in an ATP-dependent fashion. Whereas depletion of DFCP1 has a minor effect on bulk autophagic flux, DFCP1 is required to maintain the autophagic flux of p62 under both fed and starved conditions, and this is dependent on its ability to bind and hydrolyse ATP. While DFCP1 mutants defective in ATP binding or hydrolysis localize to forming omegasomes, these omegasomes fail to constrict properly in a size-dependent manner. Consequently, the release of nascent autophagosomes from large omegasomes is markedly delayed. While knockout of DFCP1 does not affect bulk autophagy, it inhibits selective autophagy, including aggrephagy, mitophagy and micronucleophagy. We conclude that DFCP1 mediates ATPase-driven constriction of large omegasomes to release autophagosomes for selective autophagy.

DOI: https://doi.org/10.1038/s41467-023-39641-9
J Phys Chem B. 2023 Jul 11.

Raman Imaging Reveals Insights into Membrane Phase Biophysics in Cells.

Yihui Shen, Lu Wei, Wei Min.

Cellular membranes are essential components of all living organisms. They are composed of a complex mixture of lipids with diverse chemical structures and crucial biological functions. The dynamic and heterogeneous nature of cellular membranes presents a challenge for studying their biophysical properties and organization in vivo. Raman imaging, particularly coherent Raman scattering techniques such as stimulated Raman scattering (SRS) microscopy, have emerged as powerful tools for studying cellular membranes with high spatial and temporal resolution and minimal perturbation. In this Review, we discuss the scientific importance and technical challenges of characterizing membrane composition in cellular contexts and how the advances of Raman imaging can provide unique insights into membrane phase behavior and organization. We also highlight recent applications of Raman imaging in studying cellular membranes and implications in diseases. In particular, the discovery of phase separation and a solid-phase intracellular membrane on endoplasmic reticulum is reviewed in detail, shedding light on the biology of lipotoxicity.

DOI: https://doi.org/10.1021/acs.jpcb.3c03125
Front Oncol. 2023 ;13 1191332

You complete me: tumor cell-myeloid cell nuclear fusion as a facilitator of organ-specific metastasis.

Alyssa J Cozzo, Michael F Coleman, Stephen D Hursting.

  Every cancer genome is unique, resulting in potentially near infinite cancer cell phenotypes and an inability to predict clinical outcomes in most cases. Despite this profound genomic heterogeneity, many cancer types and subtypes display a non-random distribution of metastasis to distant organs, a phenomenon known as organotropism. Proposed factors in metastatic organotropism include hematogenous versus lymphatic dissemination, the circulation pattern of the tissue of origin, tumor-intrinsic factors, compatibility with established organ-specific niches, long-range induction of premetastatic niche formation, and so-called "prometastatic niches" that facilitate successful colonization of the secondary site following extravasation. To successfully complete the steps required for distant metastasis, cancer cells must evade immunosurveillance and survive in multiple new and hostile environments. Despite substantial advances in our understanding of the biology underlying malignancy, many of the mechanisms used by cancer cells to survive the metastatic journey remain a mystery. This review synthesizes the rapidly growing body of literature demonstrating the relevance of an unusual cell type known as "fusion hybrid" cells to many of the hallmarks of cancer, including tumor heterogeneity, metastatic conversion, survival in circulation, and metastatic organotropism. Whereas the concept of fusion between tumor cells and blood cells was initially proposed over a century ago, only recently have technological advancements allowed for detection of cells containing components of both immune and neoplastic cells within primary and metastatic lesions as well as among circulating malignant cells. Specifically, heterotypic fusion of cancer cells with monocytes and macrophages results in a highly heterogeneous population of hybrid daughter cells with enhanced malignant potential. Proposed mechanisms behind these findings include rapid, massive genome rearrangement during nuclear fusion and/or acquisition of monocyte/macrophage features such as migratory and invasive capability, immune privilege, immune cell trafficking and homing, and others. Rapid acquisition of these cellular traits may increase the likelihood of both escape from the primary tumor site and extravasation of hybrid cells at a secondary location that is amenable to colonization by that particular hybrid phenotype, providing a partial explanation for the patterns observed in some cancers with regard to sites of distant metastases.

Keywords:  cancer metastasis; fusion hybrid; immune suppression; malignant transformation; organotropism

DOI:  https://doi.org/10.3389/fonc.2023.1191332
Cancer Discov. 2023 Jul 14. OF1-OF18

Spectrum of Response to Platinum and PARP Inhibitors in Germline BRCA-Associated Pancreatic Cancer in the Clinical and Preclinical Setting.

Chani Stossel, Maria Raitses-Gurevich, Dikla Atias, Tamar Beller, Yulia Glick Gorman, Sharon Halperin, Eyal Peer, Robert E Denroche, Amy Zhang, Faiyaz Notta, Julie M Wilson, Grainne M O'Kane, Elina Haimov Talmoud, Nora Amison, Michael Schvimer, Seth J Salpeter, Vered Bar, Adi Zundelevich, Itay Tirosh, Rotem Tal, Gal Dinstag, Yaron Kinar, Yonatan Eliezer, Uri Ben-David, Nancy S Gavert, Ravid Straussman, Steven J Gallinger, Raanan Berger, Talia Golan.

Germline BRCA-associated pancreatic ductal adenocarcinoma (glBRCA PDAC) tumors are susceptible to platinum and PARP inhibition. The clinical outcomes of 125 patients with glBRCA PDAC were stratified based on the spectrum of response to platinum/PARP inhibition: (i) refractory [overall survival (OS) <6 months], (ii) durable response followed by acquired resistance (OS <36 months), and (iii) long-term responders (OS >36 months). Patient-derived xenografts (PDX) were generated from 25 patients with glBRCA PDAC at different clinical time points. Response to platinum/PARP inhibition in vivo and ex vivo culture (EVOC) correlated with clinical response. We deciphered the mechanisms of resistance in glBRCA PDAC and identified homologous recombination (HR) proficiency and secondary mutations restoring partial functionality as the most dominant resistant mechanism. Yet, a subset of HR-deficient (HRD) patients demonstrated clinical resistance. Their tumors displayed basal-like molecular subtype and were more aneuploid. Tumor mutational burden was high in HRD PDAC and significantly higher in tumors with secondary mutations. Anti-PD-1 attenuated tumor growth in a novel humanized glBRCA PDAC PDX model. This work demonstrates the utility of preclinical models, including EVOC, to predict the response of glBRCA PDAC to treatment, which has the potential to inform time-sensitive medical decisions.
SIGNIFICANCE: glBRCA PDAC has a favorable response to platinum/PARP inhibition. However, most patients develop resistance. Additional treatment options for this unique subpopulation are needed. We generated model systems in PDXs and an ex vivo system (EVOC) that faithfully recapitulate these specific clinical scenarios as a platform to investigate the mechanisms of resistance for further drug development.

DOI: https://doi.org/10.1158/2159-8290.CD-22-0412
Front Oncol. 2023 ;13 1152553

Fluorescence microscopy imaging of mitochondrial metabolism in cancer cells.

Monika Gooz, Eduardo N Maldonado.

  Mitochondrial metabolism is an important contributor to cancer cell survival and proliferation that coexists with enhanced glycolytic activity. Measuring mitochondrial activity is useful to characterize cancer metabolism patterns, to identify metabolic vulnerabilities and to identify new drug targets. Optical imaging, especially fluorescent microscopy, is one of the most valuable tools for studying mitochondrial bioenergetics because it provides semiquantitative and quantitative readouts as well as spatiotemporal resolution of mitochondrial metabolism. This review aims to acquaint the reader with microscopy imaging techniques currently used to determine mitochondrial membrane potential (ΔΨm), nicotinamide adenine dinucleotide (NADH), ATP and reactive oxygen species (ROS) that are major readouts of mitochondrial metabolism. We describe features, advantages, and limitations of the most used fluorescence imaging modalities: widefield, confocal and multiphoton microscopy, and fluorescent lifetime imaging (FLIM). We also discus relevant aspects of image processing. We briefly describe the role and production of NADH, NADHP, flavins and various ROS including superoxide and hydrogen peroxide and discuss how these parameters can be analyzed by fluorescent microscopy. We also explain the importance, value, and limitations of label-free autofluorescence imaging of NAD(P)H and FAD. Practical hints for the use of fluorescent probes and newly developed sensors for imaging ΔΨm, ATP and ROS are described. Overall, we provide updated information about the use of microscopy to study cancer metabolism that will be of interest to all investigators regardless of their level of expertise in the field.

Keywords:  FAD; NAD(P)H; ROS; fluorescence microscopy; mitochondrial membrane potential; mitochondrial metabolism

DOI:  https://doi.org/10.3389/fonc.2023.1152553
Aging (Albany NY). 2023 Jul 12. 15

Chemically induced reprogramming to reverse cellular aging.

Jae-Hyun Yang, Christopher A Petty, Thomas Dixon-McDougall, Maria Vina Lopez, Alexander Tyshkovskiy, Sun Maybury-Lewis, Xiao Tian, Nabilah Ibrahim, Zhili Chen, Patrick T Griffin, Matthew Arnold, Jien Li, Oswaldo A Martinez, Alexander Behn, Ryan Rogers-Hammond, Suzanne Angeli, Vadim N Gladyshev, David A Sinclair.

  A hallmark of eukaryotic aging is a loss of epigenetic information, a process that can be reversed. We have previously shown that the ectopic induction of the Yamanaka factors OCT4, SOX2, and KLF4 (OSK) in mammals can restore youthful DNA methylation patterns, transcript profiles, and tissue function, without erasing cellular identity, a process that requires active DNA demethylation. To screen for molecules that reverse cellular aging and rejuvenate human cells without altering the genome, we developed high-throughput cell-based assays that distinguish young from old and senescent cells, including transcription-based aging clocks and a real-time nucleocytoplasmic compartmentalization (NCC) assay. We identify six chemical cocktails, which, in less than a week and without compromising cellular identity, restore a youthful genome-wide transcript profile and reverse transcriptomic age. Thus, rejuvenation by age reversal can be achieved, not only by genetic, but also chemical means.

Keywords:  epigenetics; information theory of aging; rejuvenation medicine; reprogramming; small molecules

DOI:  https://doi.org/10.18632/aging.204896
EMBO J. 2023 Jul 13. e113256

Ribonucleotide synthesis by NME6 fuels mitochondrial gene expression.

Nils Grotehans, Lynn McGarry, Hendrik Nolte, Vanessa Xavier, Moritz Kroker, Álvaro Jesús Narbona-Pérez, Soni Deshwal, Patrick Giavalisco, Thomas Langer, Thomas MacVicar.

  Replication of the mitochondrial genome and expression of the genes it encodes both depend on a sufficient supply of nucleotides to mitochondria. Accordingly, dysregulated nucleotide metabolism not only destabilises the mitochondrial genome, but also affects its transcription. Here, we report that a mitochondrial nucleoside diphosphate kinase, NME6, supplies mitochondria with pyrimidine ribonucleotides that are necessary for the transcription of mitochondrial genes. Loss of NME6 function leads to the depletion of mitochondrial transcripts, as well as destabilisation of the electron transport chain and impaired oxidative phosphorylation. These deficiencies are rescued by an exogenous supply of pyrimidine ribonucleosides. Moreover, NME6 is required for the maintenance of mitochondrial DNA when the access to cytosolic pyrimidine deoxyribonucleotides is limited. Our results therefore reveal an important role for ribonucleotide salvage in mitochondrial gene expression.

Keywords:  NME6; mitochondria; mitochondrial DNA; mitochondrial transcription; nucleotide metabolism

DOI:  https://doi.org/10.15252/embj.2022113256
Oncogene. 2023 Jul 11.

Pancreatic ductal adenocarcinoma induces neural injury that promotes a transcriptomic and functional repair signature by peripheral neuroglia.

Jonathan Weitz, Bharti Garg, Alexei Martsinkovskiy, Sandip Patel, Herve Tiriac, Andrew M Lowy.

Perineural invasion (PNI) is the phenomenon whereby cancer cells invade the space surrounding nerves. PNI occurs frequently in epithelial malignancies, but is especially characteristic of pancreatic ductal adenocarcinoma (PDAC). The presence of PNI portends an increased incidence of local recurrence, metastasis and poorer overall survival. While interactions between tumor cells and nerves have been investigated, the etiology and initiating cues for PNI development is not well understood. Here, we used digital spatial profiling to reveal changes in the transcriptome and to allow for a functional analysis of neural-supportive cell types present within the tumor-nerve microenvironment of PDAC during PNI. We found that hypertrophic tumor-associated nerves within PDAC express transcriptomic signals of nerve damage including programmed cell death, Schwann cell proliferation signaling pathways, as well as macrophage clearance of apoptotic cell debris by phagocytosis. Moreover, we identified that neural hypertrophic regions have increased local neuroglial cell proliferation which was tracked using EdU tumor labeling in KPC mice, as well as frequent TUNEL positivity, suggestive of a high turnover rate. Functional calcium imaging studies using human PDAC organotypic slices confirmed nerve bundles had neuronal activity, as well as contained NGFR+ cells with high sustained calcium levels, which are indicative of apoptosis. This study reveals a common gene expression pattern that characterizes solid tumor-induced damage to local nerves. These data provide new insights into the pathobiology of the tumor-nerve microenvironment during PDAC as well as other gastrointestinal cancers.

DOI: https://doi.org/10.1038/s41388-023-02775-7
Cancer Metastasis Rev. 2023 Jul 13.

How circulating tumor cluster biology contributes to the metastatic cascade: from invasion to dissemination and dormancy.

Mostafa M Nasr, Conor C Lynch.

  Circulating tumor cells (CTCs) are known to be prognostic for metastatic relapse and are detected in patients as solitary cells or cell clusters. Circulating tumor cell clusters (CTC clusters) have been observed clinically for decades and are of significantly higher metastatic potential compared to solitary CTCs. Recent studies suggest distinct differences in CTC cluster biology regarding invasion and survival in circulation. However, differences regarding dissemination, dormancy, and reawakening require more investigations compared to solitary CTCs. Here, we review the current state of CTC cluster research and consider their clinical significance. In addition, we discuss the concept of collective invasion by CTC clusters and molecular evidence as to how cluster survival in circulation compares to that of solitary CTCs. Molecular differences between solitary and clustered CTCs during dormancy and reawakening programs will also be discussed. We also highlight future directions to advance our current understanding of CTC cluster biology.

Keywords:  Cancer dissemination; Cancer dormancy; Circulating tumor cells; Circulating tumor clusters; Metastasis; Tumor microenvironment

DOI:  https://doi.org/10.1007/s10555-023-10124-z
Gastro Hep Adv. 2023 ;2(4): 532-543

Transcriptional Profile of Human Pancreatic Acinar Ductal Metaplasia.

Jinmai Jiang, Hesamedin Hakimjavadi, Julie K Bray, Corey Perkins, Alyssa Gosling, Lais daSilva, Gamze Bulut, Jamel Ali, V Wendy Setiawan, Martha Campbell-Thompson, Srikar Chamala, Thomas D Schmittgen.

   BACKGROUND AND AIMS: Aberrant acinar to ductal metaplasia (ADM), one of the earliest events involved in exocrine pancreatic cancer development, is typically studied using pancreata from genetically engineered mouse models.
METHODS: We used primary, human pancreatic acinar cells from organ donors to evaluate the transcriptional and pathway profiles during the course of ADM.
RESULTS: Following 6 days of three-dimensional culture on Matrigel, acinar cells underwent morphological and molecular changes indicative of ADM. mRNA from 14 donors' paired cells (day 0, acinar phenotype and day 6, ductal phenotype) was subjected to whole transcriptome sequencing. Acinar cell specific genes were significantly downregulated in the samples from the day 6 cultures while ductal cell-specific genes were upregulated. Several regulons of ADM were identified including transcription factors with reduced activity (PTF1A, RBPJL, and BHLHA15) and those ductal and progenitor transcription factors with increased activity (HNF1B, SOX11, and SOX4). Cells with the ductal phenotype contained higher expression of genes increased in pancreatic cancer while cells with an acinar phenotype had lower expression of cancer-associated genes.
CONCLUSION: Our findings support the relevancy of human in vitro models to study pancreas cancer pathogenesis and exocrine cell plasticity.

Keywords:  Acinar ductal metaplasia; Organoid; Pancreas; Pancreatic cancer

DOI:  https://doi.org/10.1016/j.gastha.2023.02.003
J Cachexia Sarcopenia Muscle. 2023 Jul 13.

Corylifol A ameliorates muscle atrophy by inhibiting TAOK1/p38-MAPK/FoxO3 pathway in cancer cachexia.

Ruiqin Zhang, Qiang Shen, Yueping Wang, Xue Deng, Jialing Fan, Xiaofan Gu, Meng Fan, Kun Wei, Chun-Ru Cheng, Wei-Dong Zhang, Xiong-Wen Zhang, Xuan Liu.

   BACKGROUND: Corylifol A (CYA) is one of the main active components of Psoralea corylifolia L. CYA had been reported to have ameliorating effects on dexamethasone-induced atrophy of C2C12 mouse skeletal myotubes, but its effects on cancer cachexia were unclear. Here, we checked the influence of CYA on muscle atrophy in cancer cachexia mice and tried to clarify its mechanisms.
METHODS: C26 tumour-bearing mice were applied as the animal model to examine the effects of CYA in attenuating cachexia symptoms. The in vitro cell models of TNF-α-induced C2C12 myotubes or ad-mRFP-GFP-LC3B-transfected C2C12 myotubes were used to check the influence of CYA on myotube atrophy based on both ubiquitin proteasome system (UPS) and autophagy-lysosome system. The possible direct targets of CYA were searched using the biotin-streptavidin pull-down assay and then confirmed using the Microscale thermophoresis binding assay. The levels of related signal proteins in both in vitro and in vivo experiments were examined using western blotting and immunocytochemical assay.
RESULTS: The administration of CYA prevented body weight loss and muscle wasting in C26 tumour-bearing mice without affecting tumour growth. At the end of the experiment, the body weight of mice treated with 30 mg/kg of CYA (23.59 ± 0.94 g) was significantly higher than that of the C26 model group (21.66 ± 0.56 g) with P < 0.05. The values of gastrocnemius muscle weight/body weight of mice treated with 15 or 30 mg/kg CYA (0.53 ± 0.02% and 0.54 ± 0.01%, respectively) were both significantly higher than that of the C26 model group (0.45 ± 0.01%) with P < 0.01. CYA decreased both UPS-mediated protein degradation and autophagy in muscle tissues of C26 tumour-bearing mice as well as in C2C12 myotubes treated with TNF-α. The thousand-and-one amino acid kinase 1 (TAOK1) was found to be the direct binding target of CYA. CYA inhibited the activation of TAOK1 and its downstream p38-MAPK pathway thus decreased the level and nuclear location of FoxO3. siRNA knockdown of TAOK1 or regulation of the p38-MAPK pathway using activator or inhibitor could affect the ameliorating effects of CYA on myotube atrophy.
CONCLUSIONS: CYA ameliorates cancer cachexia muscle atrophy by decreasing both UPS degradation and autophagy. The ameliorating effects of CYA on muscle atrophy might be based on its binding with TAOK1 and inhibiting the TAOK1/p38-MAPK/FoxO3 pathway.

Keywords:  Autophagy; Cancer cachexia; Corylifol A; Muscle atrophy; Proteasome

DOI:  https://doi.org/10.1002/jcsm.13288
Histochem Cell Biol. 2023 Jul 10.

OME-Zarr: a cloud-optimized bioimaging file format with international community support.

Josh Moore, Daniela Basurto-Lozada, Sébastien Besson, John Bogovic, Jordão Bragantini, Eva M Brown, Jean-Marie Burel, Xavier Casas Moreno, Gustavo de Medeiros, Erin E Diel, David Gault, Satrajit S Ghosh, Ilan Gold, Yaroslav O Halchenko, Matthew Hartley, Dave Horsfall, Mark S Keller, Mark Kittisopikul, Gabor Kovacs, Aybüke Küpcü Yoldaş, Koji Kyoda, Albane le Tournoulx de la Villegeorges, Tong Li, Prisca Liberali, Dominik Lindner, Melissa Linkert, Joel Lüthi, Jeremy Maitin-Shepard, Trevor Manz, Luca Marconato, Matthew McCormick, Merlin Lange, Khaled Mohamed, William Moore, Nils Norlin, Wei Ouyang, Bugra Özdemir, Giovanni Palla, Constantin Pape, Lucas Pelkmans, Tobias Pietzsch, Stephan Preibisch, Martin Prete, Norman Rzepka, Sameeul Samee, Nicholas Schaub, Hythem Sidky, Ahmet Can Solak, David R Stirling, Jonathan Striebel, Christian Tischer, Daniel Toloudis, Isaac Virshup, Petr Walczysko, Alan M Watson, Erin Weisbart, Frances Wong, Kevin A Yamauchi, Omer Bayraktar, Beth A Cimini, Nils Gehlenborg, Muzlifah Haniffa, Nathan Hotaling, Shuichi Onami, Loic A Royer, Stephan Saalfeld, Oliver Stegle, Fabian J Theis, Jason R Swedlow.

  A growing community is constructing a next-generation file format (NGFF) for bioimaging to overcome problems of scalability and heterogeneity. Organized by the Open Microscopy Environment (OME), individuals and institutes across diverse modalities facing these problems have designed a format specification process (OME-NGFF) to address these needs. This paper brings together a wide range of those community members to describe the cloud-optimized format itself-OME-Zarr-along with tools and data resources available today to increase FAIR access and remove barriers in the scientific process. The current momentum offers an opportunity to unify a key component of the bioimaging domain-the file format that underlies so many personal, institutional, and global data management and analysis tasks.

Keywords:  Bioimaging; Cloud; Community; Data; FAIR; Format

DOI:  https://doi.org/10.1007/s00418-023-02209-1
STAR Protoc. 2023 Jul 11. pii: S2666-1667(23)00411-2. [Epub ahead of print]4(3): 102444

Protocol for evaluation of tumor-derived exosome-induced cancer cell metastasis in a mouse model.

Yu Jin Lee, Kyeong Jin Shin, Young Chan Chae.

  Exosomes mediate intracellular communication between cancer cells and the local/distant microenvironment, which promotes systemic dissemination of cancer. Here, we present a protocol for tumor-derived exosome isolation and in vivo metastasis evaluation in a mouse model. We describe steps for isolating and characterizing exosomes, establishing a metastatic mouse model, and injecting exosomes into mouse. We then detail hematoxylin and eosin staining and analysis. This protocol can be used to investigate exosome function and identify unexplored metastatic regulators related to exosome biogenesis. For complete details on the use and execution of this protocol, please refer to Lee et al. (2023).1.

Keywords:  Cancer; Cell Biology; Model Organisms

DOI:  https://doi.org/10.1016/j.xpro.2023.102444
bioRxiv. 2023 Jul 02. pii: 2023.07.01.545937. [Epub ahead of print]

Small extracellular vesicles promote stiffness-mediated metastasis.

Alexandra Sneider, Ying Liu, Bartholomew Starich, Wenxuan Du, Carolyn Marar, Najwa Faqih, Gabrielle E Ciotti, Joo Ho Kim, Sejal Krishnan, Salma Ibrahim, Muna Igboko, Alexus Locke, Daniel M Lewis, Hanna Hong, Michelle Karl, Raghav Vij, Gabriella C Russo, Praful Nair, Estibaliz Gómez-de-Mariscal, Mehran Habibi, Arrate Muñoz-Barrutia, Luo Gu, T S Karin Eisinger-Mathason, Denis Wirtz.

Tissue stiffness is a critical prognostic factor in breast cancer and is associated with metastatic progression. Here we show an alternative and complementary hypothesis of tumor progression whereby physiological matrix stiffness affects the quantity and protein cargo of small EVs produced by cancer cells, which in turn drive their metastasis. Primary patient breast tissue produces significantly more EVs from stiff tumor tissue than soft tumor adjacent tissue. EVs released by cancer cells on matrices that model human breast tumors (25 kPa; stiff EVs) feature increased adhesion molecule presentation (ITGα 2 β 1 , ITGα 6 β 4 , ITGα 6 β 1 , CD44) compared to EVs from softer normal tissue (0.5 kPa; soft EVs), which facilitates their binding to extracellular matrix (ECM) protein collagen IV, and a 3-fold increase in homing ability to distant organs in mice. In a zebrafish xenograft model, stiff EVs aid cancer cell dissemination through enhanced chemotaxis. Moreover, normal, resident lung fibroblasts treated with stiff and soft EVs change their gene expression profiles to adopt a cancer associated fibroblast (CAF) phenotype. These findings show that EV quantity, cargo, and function depend heavily on the mechanical properties of the extracellular microenvironment.

DOI: https://doi.org/10.1101/2023.07.01.545937
Autophagy. 2023 Jul 13. 1-17

Tethering ATG16L1 or LC3 induces targeted autophagic degradation of protein aggregates and mitochondria.

Ligang Mei, Xiaorong Chen, Fujing Wei, Xue Huang, Lu Liu, Jia Yao, Jing Chen, Xunguang Luo, Zhuolin Wang, Aimin Yang.

  Proteolysis-targeting chimeras (PROTACs) based on the ubiquitin-proteasome system have made great progress in the field of drug discovery. There is mounting evidence that the accumulation of aggregation-prone proteins or malfunctioning organelles is associated with the occurrence of various age-related neurodegenerative disorders and cancers. However, PROTACs are inefficient for the degradation of such large targets due to the narrow entrance channel of the proteasome. Macroautophagy (hereafter referred to as autophagy) is known as a self-degradative process involved in the degradation of bulk cytoplasmic components or specific cargoes that are sequestered into autophagosomes. In the present study, we report the development of a generalizable strategy for the targeted degradation of large targets. Our results suggested that tethering large target models to phagophore-associated ATG16L1 or LC3 induced targeted autophagic degradation of the large target models. Furthermore, we successfully applied this autophagy-targeting degradation strategy to the targeted degradation of HTT65Q aggregates and mitochondria. Specifically, chimeras consisting of polyQ-binding peptide 1 (QBP) and ATG16L1-binding peptide (ABP) or LC3-interacting region (LIR) induced targeted autophagic degradation of pathogenic HTT65Q aggregates; and the chimeras consisting of mitochondria-targeting sequence (MTS) and ABP or LIR promoted targeted autophagic degradation of dysfunctional mitochondria, hence ameliorating mitochondrial dysfunction in a Parkinson disease cell model and protecting cells from apoptosis induced by the mitochondrial stress agent FCCP. Therefore, this study provides a new strategy for the selective proteolysis of large targets and enrich the toolkit for autophagy-targeting degradation.Abbreviations: ABP: ATG16L1-binding peptide; ATG16L1: autophagy related 16 like 1; ATTEC: autophagy-tethering compound; AUTAC: autophagy-targeting chimera; AUTOTAC: autophagy-targeting chimera; Baf A1: bafilomycin A1; BCL2: BCL2 apoptosis regulator; CALCOCO2/NDP52: calcium binding and coiled-coil domain 2; CASP3: caspase 3; CPP: cell-penetrating peptide; CQ: chloroquine phosphate; DAPI: 4',6-diamidino-2-phenylindole; DCM: dichloromethane; DMF: N,N-dimethylformamide; DMSO: dimethyl sulfoxide; EBSS: Earle's balanced salt solution; FCCP: carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone; FITC: fluorescein-5-isothiocyanate; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; GFP: green fluorescent protein; HEK293: human embryonic kidney 293; HEK293T: human embryonic kidney 293T; HPLC: high-performance liquid chromatography; HRP: horseradish peroxidase; HTT: huntingtin; LIR: LC3-interacting region; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; MFF: mitochondrial fission factor; MTS: mitochondria-targeting sequence; NBR1: NBR1 autophagy cargo receptor; NLRX1: NLR family member X1; OPTN: optineurin; P2A: self-cleaving 2A peptide; PB1: Phox and Bem1p; PBS: phosphate-buffered saline; PE: phosphatidylethanolamine; PINK1: PTEN induced kinase 1; PRKN: parkin RBR E3 ubiquitin protein ligase; PROTACs: proteolysis-targeting chimeras; QBP: polyQ-binding peptide 1; SBP: streptavidin-binding peptide; SDS-PAGE: sodium dodecyl sulfate-polyacrylamide gel electrophoresis; SPATA33: spermatogenesis associated 33; TIMM23: translocase of inner mitochondrial membrane 23; TMEM59: transmembrane protein 59; TOMM20: translocase of outer mitochondrial membrane 20; UBA: ubiquitin-associated; WT: wild type.

Keywords:  ATG16L1; HTT65Q aggregates; LC3; autophagy-targeting degradation; mitochondria; proteolysis-targeting chimeras

DOI:  https://doi.org/10.1080/15548627.2023.2234797
Clin Cancer Res. 2023 Jul 11. pii: CCR-23-0825. [Epub ahead of print]

A Novel HOXA10-Associated 5 Gene-Based Prognostic Signature for Stratification of Short-Term Survivors of Pancreatic Ductal Adenocarcinoma.

Sophia G Kisling, Pranita Atri, Ashu Shah, Jesse L Cox, Sunandini Sharma, Lynette M Smith, Dario Ghersi, Surinder K Batra.

PURPOSE: Despite the significant association of molecular subtypes with poor prognosis in pancreatic ductal adenocarcinoma (PDAC) patients, little effort has been made to identify the underlying pathway(s) responsible for this prognosis. Identifying a clinically relevant prognosis-based gene signature may be the key to improving patient outcomes.
EXPERIMENTAL DESIGN: We analyzed the transcriptomic profiles of treatment-naïve surgically resected short- and long-term survivor tumors (GSE62452) for expression and survival, followed by validation in several datasets. These results were corroborated by immunohistochemical analysis of PDAC-resected short- and long-term survivor tumors. The mechanism of this differential survival was investigated using CIBERSORT and pathway analyses.
RESULTS: We identified a short-surviving prognostic subtype of PDAC with a high degree of significance (p=0.018). One hundred thirty genes in this novel subtype were found to be regulated by a master regulator, HOXA10, and a five-gene signature derived from these genes, including BANF1, EIF4G1, MRPS10, PDIA4, and TYMS, exhibited differential expression in short-term survivors (STS) and a strong association with poor survival. This signature was further associated with the proportion of T-cells and macrophages found in STS and long-term survivors (LTS), demonstrating a potential role in PDAC immunosuppression. Pathway analyses corroborated these findings, revealing that this HOXA10-driven signature is associated with immune suppression and enhanced tumorigenesis.
CONCLUSIONS: Overall, these findings reveal the presence of a HOXA10-associated prognostic subtype that can be used to differentiate between STS and LTS patients of PDAC and inform on the molecular interactions that play a role in this poor prognosis.

DOI: https://doi.org/10.1158/1078-0432.CCR-23-0825
Cell. 2023 Jul 10. pii: S0092-8674(23)00689-X. [Epub ahead of print]

Engineering RNA export for measurement and manipulation of living cells.

Felix Horns, Joe A Martinez, Chengcheng Fan, Mehernaz Haque, James M Linton, Victoria Tobin, Leah Santat, Ailiena O Maggiolo, Pamela J Bjorkman, Carlos Lois, Michael B Elowitz.

  A system for programmable export of RNA molecules from living cells would enable both non-destructive monitoring of cell dynamics and engineering of cells capable of delivering executable RNA programs to other cells. We developed genetically encoded cellular RNA exporters, inspired by viruses, that efficiently package and secrete cargo RNA molecules from mammalian cells within protective nanoparticles. Exporting and sequencing RNA barcodes enabled non-destructive monitoring of cell population dynamics with clonal resolution. Further, by incorporating fusogens into the nanoparticles, we demonstrated the delivery, expression, and functional activity of exported mRNA in recipient cells. We term these systems COURIER (controlled output and uptake of RNA for interrogation, expression, and regulation). COURIER enables measurement of cell dynamics and establishes a foundation for hybrid cell and gene therapies based on cell-to-cell delivery of RNA.

Keywords:  RNA; delivery; export; extracellular vesicles; monitoring; non-destructive; reporter; virus-like particles

DOI:  https://doi.org/10.1016/j.cell.2023.06.013
Sci Adv. 2023 Jul 14. 9(28): eadf9915

Genome-wide CRISPR screens define determinants of epithelial-mesenchymal transition mediated immune evasion by pancreatic cancer cells.

Yuanzhuo Gu, Zhengkui Zhang, Marcel G M Camps, Ferry Ossendorp, Ruud H Wijdeven, Peter Ten Dijke.

The genetic circuits that allow cancer cells to evade immune killing via epithelial mesenchymal plasticity remain poorly understood. Here, we showed that mesenchymal-like (Mes) KPC3 pancreatic cancer cells were more resistant to cytotoxic T lymphocyte (CTL)-mediated killing than the parental epithelial-like (Epi) cells and used parallel genome-wide CRISPR screens to assess the molecular underpinnings of this difference. Core CTL-evasion genes (such as IFN-γ pathway components) were clearly evident in both types. Moreover, we identified and validated multiple Mes-specific regulators of cytotoxicity, such as Egfr and Mfge8. Both genes were significantly higher expressed in Mes cancer cells, and their depletion sensitized Mes cancer cells to CTL-mediated killing. Notably, Mes cancer cells secreted more Mfge8 to inhibit proliferation of CD8+ T cells and production of IFN-γ and TNFα. Clinically, increased Egfr and Mfge8 expression was correlated with a worse prognosis. Thus, Mes cancer cells use Egfr-mediated intrinsic and Mfge8-mediated extrinsic mechanisms to facilitate immune escape from CD8+ T cells.

DOI: https://doi.org/10.1126/sciadv.adf9915
Trends Immunol. 2023 Jul 07. pii: S1471-4906(23)00110-2. [Epub ahead of print]

Physiological role of cytokines in the regulation of mammalian metabolism.

Axel de Baat, Beckey Trinh, Helga Ellingsgaard, Marc Y Donath.

  The innate cytokine system is involved in the response to excessive food intake. In this review, we highlight recent advances in our understanding of the physiological role of three prominent cytokines, interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF), in mammalian metabolic regulation. This recent research highlights the pleiotropic and context-dependent functions in the immune-metabolic interplay. IL-1β is activated in response to overloaded mitochondrial metabolism, stimulates insulin secretion, and allocates energy to immune cells. IL-6 is released by contracting skeletal muscle and adipose tissue and directs energy from storing tissues to consuming tissues. TNF induces insulin resistance and prevents ketogenesis. Additionally, the therapeutic potential of modulating the activity of each cytokine is discussed.

Keywords:  cytokine; diabetes; humans; insulin; metabolism; obesity

DOI:  https://doi.org/10.1016/j.it.2023.06.002
J Clin Med. 2023 Jun 27. pii: 4318. [Epub ahead of print]12(13):

Modifiable Pancreatic Ductal Adenocarcinoma (PDAC) Risk Factors.

Natalia Michalak, Ewa Małecka-Wojciesko.

  This study aims to summarize the modifiable risk factors for pancreatic ductal adenocarcinoma (PDAC) that have been known for a long time, as well as information from the most recent reports. As a cancer with a late diagnosis and poor prognosis, accurate analysis of PDAC risk factors is warranted. The incidence of this cancer continues to rise, and the five-year survival rate is the lowest with respect to other tumors. The influence of cigarette smoking, alcohol consumption, and chronic pancreatitis in increasing the risk of pancreatic ductal adenocarcinoma is continually being confirmed. There are also newly emerging reports relating to the impact of lifestyle, including physical activity, the gut and oral microbiome, and hepatotropic viruses. A precise understanding of PDAC risk factors can help to identify groups of high-risk patients, and this may contribute to population awareness and education as well as earlier diagnoses with possible better treatment outcomes.

Keywords:  PDAC; pancreatic cancer; pancreatic ductal adenocarcinoma; risk factors

DOI:  https://doi.org/10.3390/jcm12134318
Nat Commun. 2023 Jul 14. 14(1): 4221

Oxidative phosphorylation is a metabolic vulnerability of endocrine therapy and palbociclib resistant metastatic breast cancers.

Rania El-Botty, Ludivine Morriset, Elodie Montaudon, Zakia Tariq, Anne Schnitzler, Marina Bacci, Nicla Lorito, Laura Sourd, Léa Huguet, Ahmed Dahmani, Pierre Painsec, Heloise Derrien, Sophie Vacher, Julien Masliah-Planchon, Virginie Raynal, Sylvain Baulande, Thibaut Larcher, Anne Vincent-Salomon, Guillaume Dutertre, Paul Cottu, Géraldine Gentric, Fatima Mechta-Grigoriou, Scott Hutton, Keltouma Driouch, Ivan Bièche, Andrea Morandi, Elisabetta Marangoni.

Resistance to endocrine treatments and CDK4/6 inhibitors is considered a near-inevitability in most patients with estrogen receptor positive breast cancers (ER + BC). By genomic and metabolomics analyses of patients' tumours, metastasis-derived patient-derived xenografts (PDX) and isogenic cell lines we demonstrate that a fraction of metastatic ER + BC is highly reliant on oxidative phosphorylation (OXPHOS). Treatment by the OXPHOS inhibitor IACS-010759 strongly inhibits tumour growth in multiple endocrine and palbociclib resistant PDX. Mutations in the PIK3CA/AKT1 genes are significantly associated with response to IACS-010759. At the metabolic level, in vivo response to IACS-010759 is associated with decreased levels of metabolites of the glutathione, glycogen and pentose phosphate pathways in treated tumours. In vitro, endocrine and palbociclib resistant cells show increased OXPHOS dependency and increased ROS levels upon IACS-010759 treatment. Finally, in ER + BC patients, high expression of OXPHOS associated genes predict poor prognosis. In conclusion, these results identify OXPHOS as a promising target for treatment resistant ER + BC patients.

DOI: https://doi.org/10.1038/s41467-023-40022-5
Cancers (Basel). 2023 Jul 05. pii: 3500. [Epub ahead of print]15(13):

Real-Life Results of Palliative Chemotherapy in Metastatic Pancreatic Ductal Adenocarcinoma.

Bianca Varzaru, Razvan A Iacob, Adina E Croitoru, Speranta M Iacob, Cristina E Radu, Stefania M Dumitrescu, Cristian Gheorghe.

   PURPOSE: To assess the efficacy of FOLFIRINOX(FFX), gemcitabine-based regimens (GB), and gemcitabine monotherapy (Gem) in patients with metastatic pancreatic ductal adenocarcinoma (mPDAC).
METHODS: This is a retrospective study that included 83 patients with mPDAC treated with first-line chemotherapy (L1) with either FFX, GB or Gem between 2015 and 2017. Progression-free survival (PFS) for L1 and second-line chemotherapy (L2) (PFS-L1 and PFS-L2) and overall survival (OS) were estimated using the Kaplan-Meier method.
RESULTS: Median PFS-L1 for FFX, GB and Gem groups was 9 months (95% (Confidence Interval) CI 2.76-15.24), 5 months (95%CI 3.44-6.56), and 5 months (95%CI 3.76-6.24), respectively (p = 0.04). OS was 14 months (95%CI 11.16-16.85), 12 months (95%CI: 9.44-11.56), and 7 months (95%CI: 5.7-8.3) for patients treated with FFX, GB, and Gem, respectively (p = 0.0001). ECOG-PS (0/1) (Hazard Ratio (HR) 6.74, p = 0.002), age > 70 years (HR 0.25, p = 0.04), body tumors (HR 2.8, p = 0.048), CA19-9 > 39 U/mL (HR 0.26, p = 0.02), and neutrophil-to-lymphocyte ratio (NLR) > 4.15 (HR 6.76, p = 0.001) were independent prognostic factors for PFS-L1. Male gender (HR 3.02, p = 0.026), ECOG-PS (0/1) (HR 4.21, p = 0.003), L1 with FFX (HR 0.255, p = 0.007), and NLR > 4.15 (HR 2.65, p = 0.04) were independent prognostic factors of OS. PFS-L2 (HR 6.91, p = 0.013) and OS-L2 (HR 6.95, p = 0.037) were significantly higher in patients first treated with FFX.
CONCLUSIONS: The OS of patients who receive FFX or GB is comparable. The best PFS-L1 belongs to the FFX group. Male gender, ECOG-PS 0/1, the FFX regimen, and NLR > 4.15 were independent predictors of OS. PFS-L2 and OS-L2 were favorably impacted by L1 with FFX.

Keywords:  metastatic pancreatic adenocarcinoma; overall survival; palliative chemotherapy; progression-free survival

DOI:  https://doi.org/10.3390/cancers15133500
Mol Oncol. 2023 Jul 15.

The stress sensor GCN2 differentially controls ribosome biogenesis in colon cancer according to the nutritional context.

Marie Piecyk, Mouna Triki, Pierre-Alexandre Laval, Cedric Duret, Joelle Fauvre, Laura Cussonneau, Christelle Machon, Jerôme Guitton, Nicolas Rama, Benjamin Gibert, Gabriel Ichim, Frederic Catez, Fleur Bourdelais, Sebastien Durand, Jean-Jacques Diaz, Isabelle Coste, Toufic Renno, Serge N Manié, Nicolas Aznar, Stephane Ansieau, Carole Ferraro-Peyret, Cedric Chaveroux.

  Nutrient availability is a key determinant of tumor cell behavior. While nutrient-rich conditions favor proliferation and tumor growth, scarcity, and particularly glutamine starvation, promotes cell dedifferentiation and chemoresistance. Here, linking ribosome biogenesis plasticity with tumor cell fate, we uncover that the amino acid sensor general control non-derepressible 2 (GCN2; also known as eIF-2-alpha kinase 4) represses the expression of the precursor of ribosomal RNA (rRNA), 47S, under metabolic stress. We show that blockade of GCN2 triggers cell death by an irremediable nucleolar stress and subsequent TP53-mediated apoptosis in patient-derived models of colon adenocarcinoma (COAD). In nutrient-rich conditions, a cell-autonomous GCN2 activity supports cell proliferation by stimulating 47S rRNA transcription, independently of the canonical integrated stress response (ISR) axis. Impairment of GCN2 activity prevents nuclear translocation of methionyl-tRNA synthetase (MetRS), resulting in nucleolar stress, mTORC1 inhibition and, ultimately, autophagy induction. Inhibition of the GCN2-MetRS axis drastically improves the cytotoxicity of RNA polymerase I (RNA pol I) inhibitors, including the first-line chemotherapy oxaliplatin, on patient-derived COAD tumoroids. Our data thus reveal that GCN2 differentially controls ribosome biogenesis according to the nutritional context. Furthermore, pharmacological co-inhibition of the two GCN2 branches and RNA pol I activity may represent a valuable strategy for elimination of proliferative and metabolically-stressed COAD cells.

Keywords:  Colon cancer; GCN2; Methionyl-tRNA synthetase; Nucleolar stress; Ribosome biogenesis

DOI:  https://doi.org/10.1002/1878-0261.13491
Cancer Cell. 2023 Jul 11. pii: S1535-6108(23)00218-0. [Epub ahead of print]

A distinct stimulatory cDC1 subpopulation amplifies CD8+ T cell responses in tumors for protective anti-cancer immunity.

Philippa Meiser, Moritz A Knolle, Anna Hirschberger, Gustavo P de Almeida, Felix Bayerl, Sebastian Lacher, Anna-Marie Pedde, Sophie Flommersfeld, Julian Hönninger, Leonhard Stark, Fabian Stögbauer, Martina Anton, Markus Wirth, Dirk Wohlleber, Katja Steiger, Veit R Buchholz, Barbara Wollenberg, Christina E Zielinski, Rickmer Braren, Daniel Rueckert, Percy A Knolle, Georgios Kaissis, Jan P Böttcher.

  Type 1 conventional dendritic cells (cDC1) can support T cell responses within tumors but whether this determines protective versus ineffective anti-cancer immunity is poorly understood. Here, we use imaging-based deep learning to identify intratumoral cDC1-CD8+ T cell clustering as a unique feature of protective anti-cancer immunity. These clusters form selectively in stromal tumor regions and constitute niches in which cDC1 activate TCF1+ stem-like CD8+ T cells. We identify a distinct population of immunostimulatory CCR7neg cDC1 that produce CXCL9 to promote cluster formation and cross-present tumor antigens within these niches, which is required for intratumoral CD8+ T cell differentiation and expansion and promotes cancer immune control. Similarly, in human cancers, CCR7neg cDC1 interact with CD8+ T cells in clusters and are associated with patient survival. Our findings reveal an intratumoral phase of the anti-cancer T cell response orchestrated by tumor-residing cDC1 that determines protective versus ineffective immunity and could be exploited for cancer therapy.

Keywords:  Dendritic cells; T cells; anti-cancer immunity; cancer immunotherapy; convolutional neural networks; deep learning; immune evasion; stem-like T cells; tumor microenvironment

DOI:  https://doi.org/10.1016/j.ccell.2023.06.008
Nutrition. 2023 Jun 14. pii: S0899-9007(23)00161-2. [Epub ahead of print]114 112132

Evaluation and management of body composition changes in cancer patients.

Junjie Wang, Shanjun Tan, Luca Gianotti, Guohao Wu.

  Wasting in cancer patients has long been recognized as a condition that adversely affects cancer patients' quality of life, treatment tolerance, and oncological outcomes. Historically, this condition was mainly evaluated by changes in body weight. However, this approach is not quite accurate because body weight is the overall change of all body compartments. Conditions such as edema and ascites can mask the severity of muscle and adipose tissue depletion. Changes in body composition assessment in cancer patients have historically been underappreciated because of the limited availability of measurement tools. As more evidence highlighting the importance of body composition has emerged, it is imperative to apply a more precise evaluation of nutritional status and a more targeted approach to provide nutritional support for cancer patients. In this review, we will discuss the modalities for evaluating body composition and how to manage body composition changes in cancer patients.

Keywords:  Body composition; Cachexia; Cancer; Skeletal muscle; sarcopenia

DOI:  https://doi.org/10.1016/j.nut.2023.112132
Br J Surg. 2023 Jul 13. pii: znad210. [Epub ahead of print]

Predictive value of baseline serum carbohydrate antigen 19-9 level on treatment effect of neoadjuvant chemoradiotherapy in patients with resectable and borderline resectable pancreatic cancer in two randomized trials.

Dutch Pancreatic Cancer Group

BACKGROUND: Guidelines suggest that the serum carbohydrate antigen (CA19-9) level should be used when deciding on neoadjuvant treatment in patients with resectable and borderline resectable pancreatic ductal adenocarcinoma (hereafter referred to as pancreatic cancer). In patients with resectable pancreatic cancer, neoadjuvant therapy is advised when the CA19-9 level is 'markedly elevated'. This study investigated the impact of baseline CA19-9 concentration on the treatment effect of neoadjuvant chemoradiotherapy (CRT) in patients with resectable and borderline resectable pancreatic cancers.
METHODS: In this post hoc analysis, data were obtained from two RCTs that compared neoadjuvant CRT with upfront surgery in patients with resectable and borderline resectable pancreatic cancers. The effect of neoadjuvant treatment on overall survival was compared between patients with a serum CA19-9 level above or below 500 units/ml using the interaction test.
RESULTS: Of 296 patients, 179 were eligible for analysis, 90 in the neoadjuvant CRT group and 89 in the upfront surgery group. Neoadjuvant CRT was associated with superior overall survival (HR 0.67, 95 per cent c.i. 0.48 to 0.94; P = 0.019). Among 127 patients (70, 9 per cent) with a low CA19-9 level, median overall survival was 23.5 months with neoadjuvant CRT and 16.3 months with upfront surgery (HR 0.63, 0.42 to 0.93). For 52 patients (29 per cent) with a high CA19-9 level, median overall survival was 15.5 months with neoadjuvant CRT and 12.9 months with upfront surgery (HR 0.82, 0.45 to 1.49). The interaction test for CA19-9 level exceeding 500 units/ml on the treatment effect of neoadjuvant CRT was not significant (P = 0.501).
CONCLUSION: Baseline serum CA19-9 level defined as either high or low has prognostic value, but was not associated with the treatment effect of neoadjuvant CRT in patients with resectable and borderline resectable pancreatic cancers, in contrast with current guideline advice.

DOI: https://doi.org/10.1093/bjs/znad210
Nat Biotechnol. 2023 Jul 10.

Whole-body cellular mapping in mouse using standard IgG antibodies.

Hongcheng Mai, Jie Luo, Luciano Hoeher, Rami Al-Maskari, Izabela Horvath, Ying Chen, Florian Kofler, Marie Piraud, Johannes C Paetzold, Jennifer Modamio, Mihail Todorov, Markus Elsner, Farida Hellal, Ali Ertürk.

Whole-body imaging techniques play a vital role in exploring the interplay of physiological systems in maintaining health and driving disease. We introduce wildDISCO, a new approach for whole-body immunolabeling, optical clearing and imaging in mice, circumventing the need for transgenic reporter animals or nanobody labeling and so overcoming existing technical limitations. We identified heptakis(2,6-di-O-methyl)-β-cyclodextrin as a potent enhancer of cholesterol extraction and membrane permeabilization, enabling deep, homogeneous penetration of standard antibodies without aggregation. WildDISCO facilitates imaging of peripheral nervous systems, lymphatic vessels and immune cells in whole mice at cellular resolution by labeling diverse endogenous proteins. Additionally, we examined rare proliferating cells and the effects of biological perturbations, as demonstrated in germ-free mice. We applied wildDISCO to map tertiary lymphoid structures in the context of breast cancer, considering both primary tumor and metastases throughout the mouse body. An atlas of high-resolution images showcasing mouse nervous, lymphatic and vascular systems is accessible at http://discotechnologies.org/wildDISCO/atlas/index.php .

DOI: https://doi.org/10.1038/s41587-023-01846-0