bims-medica 2023-10-08 papers

bims-medica

Biomed News

on Metabolism and diet in cancer

Issue of 2023–10–08
twenty papers selected by
Brett Chrest, East Carolina University

Sex-specific differences in metabolic hormone and adipose tissue dynamics induced by moderate low-carbohydrate and ketogenic diet.
Exploiting Enhanced Lipid Metabolism in Glioblastoma through Diet Modification.
Editorial: Cancer metabolism: molecular insights, metabolic crosstalk in the tumor microenvironment, and implications for therapy.
Portable optical spectroscopic assay for non-destructive measurement of key metabolic parameters on in vitro cancer cells and organotypic fresh tumor slices.
Nuclear VCP drives colorectal cancer progression by promoting fatty acid oxidation.
Promoting the Anti-Tumor Activity of Radiotherapy on Lung Cancer through a Modified Ketogenic Diet and the AMPK Signaling Pathway.
Application of fluorescence lifetime imaging microscopy to monitor glucose metabolism in pancreatic islets in vivo.
Obesity control and liver health in breast cancer: Normalized hepatic elasticity after ketogenic diet.
Intestinal-specific Hdac3 deletion increases susceptibility to colitis and small intestinal tumour development in mice fed a high fat diet.
Rat models of diet-induced obesity and metabolic dysregulation: Current trends, shortcomings and considerations for future research.
High-fat diet promotes tumor growth in the patient-derived orthotopic xenograft (PDOX) mouse model of ER positive endometrial cancer.
Ketogenesis promotes tolerance to Pseudomonas aeruginosa pulmonary infection.
Depleting glioblastoma cells of very long-chain acyl-CoA synthetase 3 (ACSVL3) produces metabolic alterations in non-lipid pathways.
Hypoxia induced deregulation of sphingolipids in colon cancer is a prognostic marker for patient outcome.
Establishing a Physiologic Human Vascularized Micro-Tumor Model for Cancer Research.
Multi-omics analysis revealed the mitochondrial-targeted drug combination to suppress the development of lung cancer.
Aminooxyacetic acid hemihydrochloride leads to decreased intracellular ATP levels and altered cell cycle of prostate cancer cells by suppressing energy metabolism.
Rewiring of mitochondrial metabolism in therapy-resistant cancers: permanent and plastic adaptations.
Spatiotemporal modeling of chemoresistance evolution in breast tumors uncovers dependencies on SLC38A7 and SLC46A1.
Intensified NK cell therapy in combination with low-dose chemoradiotherapy against human colorectal cancer.

Sci Rep. 2023 09 30. 13(1): 16465

Sex-specific differences in metabolic hormone and adipose tissue dynamics induced by moderate low-carbohydrate and ketogenic diet.

Ilya Smolensky, Kilian Zajac-Bakri, Timothy Sasha Odermatt, Catherine Brégère, John F Cryan, Raphael Guzman, Katharina Timper, Dragos Inta.

Low-carbohydrates diets are increasingly used to treat obesity and metabolic disorders. A very low-carbohydrate, ketogenic diet is hard to follow and, due to the very high fat content, linked to severe side effects, like hyperlipidemia and atherogenesis. Therefore, a less restrictive, unsaturated fat-based low-carbohydrate diet appears as a promising alternative. Since neither sex differences, nor their effect on specific metabolic hormones and adipose tissue compartments have been investigated thoroughly in these diets, we aimed to analyze their dynamics and metabolic factors in mice. We found a significant sexual dimorphism with decreased body weight and subcutaneous fat only in males on ketogenic diet, while diminished insulin, elevated ghrelin and FGF-21 were present with a differential time course in both sexes. The non-ketogenic moderate low-carbohydrate diet increased body weight and perigonadal fat in females, but induced leptin elevation in males. Both diets enhanced transiently TNFɑ only in males and had no impact on behavior. Altogether, these results reveal complex sex-dependent effect of dietary interventions, indicating unexpectedly females as more prone to unfavorable metabolic effects of low-carbohydrate diets.

DOI: https://doi.org/10.1038/s41598-023-43587-9
Int J Radiat Oncol Biol Phys. 2023 Oct 01. pii: S0360-3016(23)05329-4. [Epub ahead of print]117(2S): e113-e114

Exploiting Enhanced Lipid Metabolism in Glioblastoma through Diet Modification.

S Kant, Y Zhao, P Kesarwani, P Chinnaiyan.

PURPOSE/OBJECTIVE(S): Enhanced lipid metabolism has emerged as a central metabolic node in glioblastoma, serving as a 'gain of function' that allows these cells to efficiently adapt to their dynamic tumor microenvironment. Seemingly contradictory to this, pre-clinical studies have demonstrated anti-tumor activity in mice fed a high-fat/low-carbohydrate ketogenic diet (KD), both alone and in combination with radiation therapy (RT). In this study, we sought to identify mechanisms underlying the antitumor activity of a KD in glioblastoma from a metabolic perspective to better understand factors contributing to this apparent disconnect.
MATERIALS/METHODS: Immunocompromised and immunocompetent mice were injected orthotopically with human and mouse-derived glioblastoma cell lines and randomized to four treatment arms. Mice were fed ad libitum a standard diet (SD), KD (Bio-Serve), or a modified unsaturated fatty acid (uFA) rich diet (MD; 60/30/10: fat/protein/carb) alone or in combination with hypofractionated RT (6 Gy x 3). Global metabolomic profiling of tumors and serum were carried out using LC/GC-MS. Lipid droplets were analyzed by flow cytometer and confocal microscopy using BODIPY staining and free fatty acids were measured using a commercially available kit.
RESULTS: A KD demonstrated independent anti-tumor activity and potent synergy with RT in two aggressive glioblastoma models. Metabolomic profiling of tumors revealed significant changes in tumor metabolism in KD-fed mice when compared to SD, with an accumulation of uFAs being a key finding. We therefore sought to determine if this accumulation of fatty acids in KD mice contributed towards the observed anti-tumor activity. Consistent with in vivo results, in vitro studies using the uFA linoleic acid demonstrated anti-proliferative activity, reduced clonogenic capacity, and potent synergy when combined with RT in glioblastoma cells. Through a series of investigations, we went on to determine that this anti-tumor activity was attributed to the ability of uFA to override lipid storage homeostasis in glioblastoma cells, resulting in lipotoxicity. Based on these findings, we hypothesized high fat concentrations, rather than carbohydrate restriction, contributed to the anti-tumor activity of a KD. To test this, we generated a MD rich in uFA that did not require carbohydrate restriction. Similar to a KD, mice fed a MD demonstrated both independent anti-tumor activity and potent synergy when combined with RT.
CONCLUSION: High concentrations of uFA represents a key factor underlying the anti-tumor activity of a KD in glioblastoma by targeting lipid homeostasis. A MD consisting of high concentrations of uFA without carbohydrate restriction demonstrates promising anti-tumor activity in glioblastoma models. As a major limitation of a KD is tolerability, particularly in glioblastoma patients, a MD represents a promising form of dietary modification that may be translated clinically.

DOI: https://doi.org/10.1016/j.ijrobp.2023.06.895
Front Oncol. 2023 ;13 1289397

Editorial: Cancer metabolism: molecular insights, metabolic crosstalk in the tumor microenvironment, and implications for therapy.

Balkrishna Chaube, Parmanand Malvi.



Keywords:  bioenergetics; cancer; cancer/immune metabolism; metabolic disorder and cancer; metabolic imaging; metabolomics; tumor microenvironment

DOI:  https://doi.org/10.3389/fonc.2023.1289397
Biomed Opt Express. 2023 Aug 01. 14(8): 4065-4079

Portable optical spectroscopic assay for non-destructive measurement of key metabolic parameters on in vitro cancer cells and organotypic fresh tumor slices.

Jing Yan, Carlos Frederico Lima Goncalves, Madison O Korfhage, Md Zahid Hasan, Teresa W-M Fan, Xiaoqin Wang, Caigang Zhu.

To enable non-destructive metabolic characterizations on in vitro cancer cells and organotypic tumor models for therapeutic studies in an easy-to-access way, we report a highly portable optical spectroscopic assay for simultaneous measurement of glucose uptake and mitochondrial function on various cancer models with high sensitivity. Well-established breast cancer cell lines (MCF-7 and MDA-MB-231) were used to validate the optical spectroscopic assay for metabolic characterizations, while fresh tumor samples harvested from both animals and human cancer patients were used to test the feasibility of our optical metabolic assay for non-destructive measurement of key metabolic parameters on organotypic tumor slices. Our optical metabolic assay captured that MCF-7 cells had higher mitochondrial metabolism, but lower glucose uptake compared to the MDA-MB-231 cells, which is consistent with our microscopy imaging and flow cytometry data, as well as the published Seahorse Assay data. Moreover, we demonstrated that our optical assay could non-destructively measure both glucose uptake and mitochondrial metabolism on the same cancer cell samples at one time, which remains challenging by existing metabolic tools. Our pilot tests on thin fresh tumor slices showed that our optical assay captured increased metabolic activities in tumors compared to normal tissues. Our non-destructive optical metabolic assay provides a cost-effective way for future longitudinal therapeutic studies using patient-derived organotypic fresh tumor slices through the lens of tumor energetics, which will significantly advance translational cancer research.

DOI: https://doi.org/10.1364/BOE.497127
Proc Natl Acad Sci U S A. 2023 Oct 10. 120(41): e2221653120

Nuclear VCP drives colorectal cancer progression by promoting fatty acid oxidation.

Youwei Huang, Fang Wang, Xi Lin, Qing Li, Yuli Lu, Jiayu Zhang, Xi Shen, Jingyi Tan, Zixi Qin, Jiahong Chen, Xueqin Chen, Guopeng Pan, Xiangyu Wang, Yuequan Zeng, Shangqi Yang, Jun Liu, Fan Xing, Kai Li, Haipeng Zhang.

  Fatty acid oxidation (FAO) fuels many cancers. However, knowledge of pathways that drive FAO in cancer remains unclear. Here, we revealed that valosin-containing protein (VCP) upregulates FAO to promote colorectal cancer growth. Mechanistically, nuclear VCP binds to histone deacetylase 1 (HDAC1) and facilitates its degradation, thus promoting the transcription of FAO genes, including the rate-limiting enzyme carnitine palmitoyltransferase 1A (CPT1A). FAO is an alternative fuel for cancer cells in environments exhibiting limited glucose availability. We observed that a VCP inhibitor blocked the upregulation of FAO activity and CPT1A expression triggered by metformin in colorectal cancer (CRC) cells. Combined VCP inhibitor and metformin prove more effective than either agent alone in culture and in vivo. Our study illustrates the molecular mechanism underlying the regulation of FAO by nuclear VCP and demonstrates the potential therapeutic utility of VCP inhibitor and metformin combination treatment for colorectal cancer.

Keywords:  VCP; colorectal cancer; combination therapy; fatty acid oxidation; metformin

DOI:  https://doi.org/10.1073/pnas.2221653120
Int J Radiat Oncol Biol Phys. 2023 Oct 01. pii: S0360-3016(23)05666-3. [Epub ahead of print]117(2S): e268-e269

Promoting the Anti-Tumor Activity of Radiotherapy on Lung Cancer through a Modified Ketogenic Diet and the AMPK Signaling Pathway.

L Xiao, J Lv, T Li.

PURPOSE/OBJECTIVE(S): Taking advantage of the characteristics of high metabolic heterogeneity of tumor cells, the modified ketogenic diet (KD) combined with radiotherapy was used to investigate and analyze the radiosensitivity of a lung cancer model from the perspective of energy metabolism.
MATERIALS/METHODS: Different concentrations of glucose and βhydroxybutyrate (βHB) were used at the cellular level to simulate the level of ketone bodies. A cell counting kit was used to detect the effect of different concentrations of glucose (2.78mM, 5.56mM, 12.5mM, and 25mM) and βHB (0mM, 5mM, and 10mM) combined with radiotherapy on the proliferation of LLC cells. Flow cytometry was used to detect tumor cell cycle and apoptosis. Immunofluorescence was used to detect the expression of γH2AX, a DNA damage marker, and western blot was used to detect the expression of AMPK and ρ-AMPK. At the animal level, C57BL/6J female mice were used to establish a transplanted tumor model of lung cancer, and fed with different fat ratio diets combined with radiotherapy. The volume, tumor size, blood glucose level, blood ketone level, survival time and safety of the mice were monitored and observed.
RESULTS: The LLC cells were treated with different concentrations of glucose and βHB. The results showed that the survival rate of LLC cells decreased significantly with the increase of irradiation dose when the glucose concentration was 5.56mM and 2.78mM; However, the survival rate of cells in low glucose medium added with βHB was significantly lower than that of the control group, and the survival rate of LLC decreased significantly with the extension of culture time after irradiation (p < 0.001). After irradiation, LG (low glucose) group, LG+βHB 5mM group and LG+βHB 10mM group had a significantly higher proportion of G2 phase, and a significantly higher proportion of early and late phase than the control group. γH2AX foci were detected in LG group, LG +βHB 5mM group and LG +βHB 10mM group at 2h and 24h after radiotherapy, which were significantly higher than those in the control group (p < 0.05). The median survival time was 38 days in the PT group, 55 days in the PT+RT group, 41 days in the 45F group, and not reached in the 45F+RT group. HE staining showed no tumor metastasis and toxic side effects in liver and kidney. The expression of ρ-AMPK/AMPK in the combined treatment group was higher than that in the other groups. The expression of ρ-AMPK/AMPK in RT, 45F and combined treatment group was higher than that in PT group. The expression of ρ-AMPK/AMPK in RT group was higher than that in 45F group, and the difference was statistically significant (p < 0.01).
CONCLUSION: Modified ketogenic diet can enhance the anti-tumor effect of radiotherapy in LLC tumor-bearing mice by reducing glucose and increasing the energy supply ratio from fat.

DOI: https://doi.org/10.1016/j.ijrobp.2023.06.1232
Biomed Opt Express. 2023 Aug 01. 14(8): 4170-4178

Application of fluorescence lifetime imaging microscopy to monitor glucose metabolism in pancreatic islets in vivo.

Zhongying Wang, Maani Archang, Tatyana Gurlo, Elaine Wong, Scott E Fraser, Peter C Butler.

Glucose stimulated insulin secretion is mediated by glucose metabolism via oxidative phosphorylation generating ATP that triggers membrane depolarization and exocytosis of insulin. In stressed beta cells, glucose metabolism is remodeled, with enhanced glycolysis uncoupled from oxidative phosphorylation, resulting in the impaired glucose-mediated insulin secretion characteristic of diabetes. Relative changes in glycolysis and oxidative phosphorylation can be monitored in living cells using the 3-component fitting approach of fluorescence lifetime imaging microscopy (FLIM). We engrafted pancreatic islets onto the iris to permit in vivo FLIM monitoring of the trajectory of glucose metabolism. The results show increased oxidative phosphorylation of islet cells (∼90% beta cells) in response to hyperglycemia; in contrast red blood cells traversing the islets maintained exclusive glycolysis as expected in the absence of mitochondria.

DOI: https://doi.org/10.1364/BOE.493722
Heliyon. 2023 Oct;9(10): e20449

Obesity control and liver health in breast cancer: Normalized hepatic elasticity after ketogenic diet.

Aleksandar Georgiev, Lyubomir Chervenkov, Daniela Koleva, Vanya Anastasova.

  Most socially significant diseases, including breast cancer, are undeniably linked to obesity. Recently, a positive relationship between excessive weight and increased risk of breast cancer poor outcomes has been proved. Liver integrity is an essential point during chemotherapy. Consequently, a hepatic safe therapeutic approach for managing obesity in patients with breast cancer should be initiated. Our study aimed to assess the impact of the ketogenic diet on body mass index (BMI) and to evaluate its safety on liver function in female patients with breast cancer. The study comprised 520 women with ductal breast cancer who underwent a 60-day modified ketogenic diet. BMI, prothrombin time (PT), activated partial thromboplastin clotting time (aPTT), aspartate aminotransferase to platelet ratio index (APRI), and ultrasound liver elasticity was evaluated before and after the diet. The results showed a significant decrease in BMI and an improvement in ultrasound liver elasticity in all the participants after completing the diet. Before the KD, the participants' median BMI was 35.0 kg/m2, and after the 60-day diet, the median BMI was reduced to 30.0 kg/m2. No significant liver parameter changes were found after the diet. In conclusion, we can safely promote the keto diet amongst individuals with an increased chance of developing breast cancer for a better disease prevention.

Keywords:  Breast cancer; Ketogenic diet; Ketones; Weight reduction; liver safety

DOI:  https://doi.org/10.1016/j.heliyon.2023.e20449
Am J Physiol Gastrointest Liver Physiol. 2023 Oct 03.

Intestinal-specific Hdac3 deletion increases susceptibility to colitis and small intestinal tumour development in mice fed a high fat diet.

Irvin Ng, Ian Y Luk, Rebecca Nightingale, Camilla M Reehorst, Mercedes Davalos-Salas, Laura J Jenkins, Chun Fong, David S Williams, Matthew J Watt, Amardeep S Dhillon, John M Mariadason.

  High fat diets, and inflammation are risk factors for colon cancer, however the underlying mechanisms remain to be fully elucidated. The transcriptional co-repressor HDAC3 has recently emerged as a key regulator of intestinal epithelial responses to diet and inflammation with intestinal-specific Hdac3 deletion (Hdac3IKO) in mice increasing lipid oxidation genes and the rate of lipid oxidation in enterocytes. Hdac3IKO mice are also predisposed to experimentally induced colitis, however whether this is driven by the intestinal metabolic reprogramming, and whether this predisposes these mice to intestinal tumourigenesis is unknown. Herein, we examined the effects of intestinal-specific Hdac3 deletion on colitis-associated intestinal tumourigenesis in mice fed a standard (STD) or HF diet (HFD). Hdac3IKO mice were highly prone to experimentally induced colitis, which was further enhanced by a HFD. Hdac3deletion also accelerated intestinal tumour development, specifically when fed a HFD and most notably in the small intestine where lipid absorption is maximal. Expression of proteins involved in fatty acid metabolism and oxidation (SCD1, EHHADH) were elevated in the small intestine of Hdac3IKO mice fed a HFD, and these mice displayed increased levels of lipid peroxidation, DNA damage, and apoptosis in their villi, as well as extensive expansion of the stem cell and progenitor cell compartment. These findings reveal a novel role for Hdac3 in suppressing colitis and intestinal tumorigenesis, particularly in the context of consumption of a HFD, and reveal a potential mechanism by which HFDs may increase intestinal tumorigenesis by increasing lipid oxidation, DNA damage and intestinal epithelial cell turnover.

Keywords:  AOM/DSS; HDAC3; colon; high fat diet; intestine

DOI:  https://doi.org/10.1152/ajpgi.00160.2023
Obes Res Clin Pract. 2023 Oct 01. pii: S1871-403X(23)00113-8. [Epub ahead of print]

Rat models of diet-induced obesity and metabolic dysregulation: Current trends, shortcomings and considerations for future research.

Hanél Sadie-Van Gijsen, Liske Kotzé-Hörstmann.

  Rat diet-induced obesity and metabolic dysregulation (DIO/DIMD) is widely used as a pre-clinical model for human obesity and for testing weight-loss interventions. The aim of this review was to utilise a systematic literature survey of rat DIO/DIMD studies as a tool to document trends around study design and metabolic outcomes of these studies, and to consider ways in which the design of these studies may be improved to enhance the relevance thereof for human obesity research. In total, 110 comparisons between control and obesogenic dietary groups were included in the survey. Young male rats were found to be the model of choice, but fewer than 50% of studies provided comprehensive information about diet composition and energy intake. In addition, it was found that the majority of expected DIO/DIMD responses (hyperglycemia, hyperinsulinemia, dyslipidemia, hypoadiponectinemia) occurred at < 80% frequency, drawing into question the concept of a "typical" or "appropriate" response. We discuss the impact of differences in diet composition and energy intake on metabolic outcomes against the context of large heterogeneity of obesogenic diets employed in rat DIO/DIMD studies, and provide recommendations for the improvement of reporting standards around diet composition and dietary intake. In addition, we highlight the lack of data from female and older rats and describe considerations around the inclusion of sex and age as a variable in rat DIO/DIMD studies, aiming towards improving the applicability of these studies as a model of human obesity, which is most prevalent in women and older individuals.

Keywords:  Diet composition; Diet-induced obesity; Energy intake; Female rats; Metabolic outcomes; Systematic literature survey

DOI:  https://doi.org/10.1016/j.orcp.2023.09.010
Sci Rep. 2023 10 02. 13(1): 16537

High-fat diet promotes tumor growth in the patient-derived orthotopic xenograft (PDOX) mouse model of ER positive endometrial cancer.

Ke Shen, Dandan Shen, Dongdong Jin, Yichao Zheng, Yuanhang Zhu, Xinyue Zhao, Zhenan Zhang, Nannan Wang, Huanhuan Chen, Li Yang.

Endometrial cancer, one of the common gynecological malignancies, is affected by several influencing factors. This study established a unique patient-derived orthotopic xenograft (PDOX) nude mouse model for the study of influencing factors in ER positive endometrial cancer. The aim of this study was to demonstrate that a high-fat diet can affect the growth of ER positive endometrial cancer PDOX model tumors. The tumor tissues were expanded by subcutaneous transplantation in nude mice, and then the subcutaneous tumor tissues were orthotopically implanted into the nude mouse uterus to establish the PDOX model. After modeling, they were divided into high-fat diet group and normal diet group for 8 weeks of feeding, which showed that high-fat diet significantly promoted tumor growth (P < 0.001) and increased the protein expression level of ERα in tumor tissues. This study demonstrates that PDOX models of endometrial cancer can embody the role of dietary influences on tumor growth and that this model has the potential for preclinical studies of cancer promoting factors.

DOI: https://doi.org/10.1038/s41598-023-43797-1
Cell Metab. 2023 Oct 03. pii: S1550-4131(23)00332-7. [Epub ahead of print]35(10): 1767-1781.e6

Ketogenesis promotes tolerance to Pseudomonas aeruginosa pulmonary infection.

Kira L Tomlinson, Ying-Tsun Chen, Alex Junker, AndreaCarola Urso, Tania Wong Fok Lung, Danielle Ahn, Casey E Hofstaedter, Swikrity U Baskota, Robert K Ernst, Alice Prince, Sebastián A Riquelme.

  Pseudomonas aeruginosa is a common cause of pulmonary infection. As a Gram-negative pathogen, it can initiate a brisk and highly destructive inflammatory response; however, most hosts become tolerant to the bacterial burden, developing chronic infection. Using a murine model of pneumonia, we demonstrate that this shift from inflammation to disease tolerance is promoted by ketogenesis. In response to pulmonary infection, ketone bodies are generated in the liver and circulate to the lungs where they impose selection for P. aeruginosa strains unable to display surface lipopolysaccharide (LPS). Such keto-adapted LPS strains fail to activate glycolysis and tissue-damaging cytokines and, instead, facilitate mitochondrial catabolism of fats and oxidative phosphorylation (OXPHOS), which maintains airway homeostasis. Within the lung, P. aeruginosa exploits the host immunometabolite itaconate to further stimulate ketogenesis. This environment enables host-P. aeruginosa coexistence, supporting both pathoadaptive changes in the bacteria and the maintenance of respiratory integrity via OXPHOS.

Keywords:  OXPHOS; Pseudomonas aeruginosa; bioenergetics; disease tolerance; infection; inflammation; itaconate; ketogenesis; ketogenic diet; pneumonia

DOI:  https://doi.org/10.1016/j.cmet.2023.09.001
bioRxiv. 2023 Sep 18. pii: 2023.09.18.558236. [Epub ahead of print]

Depleting glioblastoma cells of very long-chain acyl-CoA synthetase 3 (ACSVL3) produces metabolic alterations in non-lipid pathways.

Elizabeth A Kolar, Xiaohai Shi, Emily M Clay, Yanqiu Liu, Shuli Xia, Cissy Zhang, Anne Le, Paul A Watkins.

Knockout (KO) of the fatty acid-activation enzyme very long-chain acyl-CoA synthetase 3 (ACSVL3; SLC27A3) in U87MG glioblastoma cells reduced their malignant growth properties both in vitro and in xenografts. These U87-KO glioma cells grew at a slower rate, became adherence-dependent, and were less invasive than parental U87 cells. U87-KO cells produced fewer, slower-growing subcutaneous and intracranial tumors when implanted in NOD-SCID mice. Thus, depleting U87MG cells of ACSVL3 restored these cells to a phenotype more like that of normal astrocytes. To understand the mechanisms underlying these beneficial changes, we investigated several possibilities, including the effects of ACSVL3 depletion on carbohydrate metabolism. Proteomic and metabolomic profiling indicated that ACSVL3 KO produced changes in glucose and energy metabolism. Even though protein levels of glucose transporters GLUT1 and GLUT3 were reduced by KO, cellular uptake of labeled 2-deoxyglucose was unaffected. Glucose oxidation to CO 2 was reduced nearly 7-fold by ACSVL3 depletion, and the cellular glucose level was 25% higher in KO cells. Glycolytic enzymes were upregulated by KO, but metabolic intermediates were essentially unchanged. Surprisingly, lactate production and the levels of lactate dehydrogenase isozymes LDHA and LDHB were elevated by ACSVL3 KO. The activity of the pentose phosphate pathway was found to be lower in KO cells. Citric acid cycle enzymes, electron transport chain complexes, and ATP synthase protein levels were all reduced by ACSVL3 depletion. Mitochondria were elongated in KO cells, but had a more punctate morphology in U87 cells. The mitochondrial potential was unaffected by lack of ACSVL3. We conclude that the beneficial effects of ACSVL3 depletion in human glioblastoma cells may result in part from alterations in diverse metabolic processes that are not directly related to role(s) of this enzyme in fatty acid and/or lipid metabolism. (Supported by NIH 5R01NS062043 and KKI institutional funds.).

DOI: https://doi.org/10.1101/2023.09.18.558236
Biochim Biophys Acta Mol Basis Dis. 2023 Oct 04. pii: S0925-4439(23)00272-7. [Epub ahead of print] 166906

Hypoxia induced deregulation of sphingolipids in colon cancer is a prognostic marker for patient outcome.

Khadija El Hindi, Sebastian Brachtendorf, Jennifer C Hartel, Christoph Renné, Kerstin Birod, Karin Schilling, Sandra Labocha, Dominique Thomas, Nerea Ferreirós, Lisa Hahnefeld, Erika Dorochow, Domenico Del Turco, Thomas Deller, Klaus Scholich, Dominik C Fuhrmann, Andreas Weigert, Bernhard Brüne, Gerd Geisslinger, Ilka Wittig, Karl-Heinrich Link, Sabine Grösch.

  Sphingolipids are important for the physicochemical properties of cellular membranes and deregulated in tumors. In human colon cancer tissue ceramide synthase (CerS) 4 and CerS5 are reduced which correlates with a reduced survival probability of late-stage colon cancer patients. Both enzymes are reduced after hypoxia in advanced colorectal cancer (CRC) cells (HCT-116, SW620) but not in non-metastatic CRC cells (SW480, Caco-2). Downregulation of CerS4 or CerS5 in advanced CRC cells enhanced tumor formation in nude mice and organoid growth in vitro. This was accompanied by an enhanced proliferation rate and metabolic changes leading to a shift towards the Warburg effect. In contrast, CerS4 or CerS5 depletion in Caco-2 cells reduced tumor growth in vivo. Lipidomic and proteomic analysis of membrane fractions revealed significant changes in tumor-promoting cellular pathways and cellular transporters. This study identifies CerS4 and CerS5 as prognostic markers for advanced colon cancer patients and provides a comprehensive overview about the associated cellular metabolic changes. We propose that the expression level of CerS4 and CerS5 in colon tumors could serve as a basis for decision-making for personalized treatment of advanced colon cancer patients. Trial registration: The study was accredited by the study board of the Deutsche Krebsgesellschaft (Registration No: St-D203, 2017/06/30, retrospectively registered).

Keywords:  Biomarker; Lipidomic; Metabolism; Oxidative consumption rate, hypoxia; Proteomic

DOI:  https://doi.org/10.1016/j.bbadis.2023.166906
J Vis Exp. 2023 09 15.

Establishing a Physiologic Human Vascularized Micro-Tumor Model for Cancer Research.

Stephanie J Hachey, Daniela Gaebler, Christopher C W Hughes.

A lack of validated cancer models that recapitulate the tumor microenvironment of solid cancers in vitro remains a significant bottleneck for preclinical cancer research and therapeutic development. To overcome this problem, we have developed the vascularized microtumor (VMT), or tumor chip, a microphysiological system that realistically models the complex human tumor microenvironment. The VMT forms de novo within a microfluidic platform by co-culture of multiple human cell types under dynamic, physiological flow conditions. This tissue-engineered micro-tumor construct incorporates a living perfused vascular network that supports the growing tumor mass just as newly formed vessels do in vivo. Importantly, drugs and immune cells must cross the endothelial layer to reach the tumor, modeling in vivo physiological barriers to therapeutic delivery and efficacy. Since the VMT platform is optically transparent, high-resolution imaging of dynamic processes such as immune cell extravasation and metastasis can be achieved with direct visualization of fluorescently labeled cells within the tissue. Further, the VMT retains in vivo tumor heterogeneity, gene expression signatures, and drug responses. Virtually any tumor type can be adapted to the platform, and primary cells from fresh surgical tissues grow and respond to drug treatment in the VMT, paving the way toward truly personalized medicine. Here, the methods for establishing the VMT and utilizing it for oncology research are outlined. This innovative approach opens new possibilities for studying tumors and drug responses, providing researchers with a powerful tool to advance cancer research.

DOI: https://doi.org/10.3791/65865
J Cancer Res Clin Oncol. 2023 Oct 02.

Multi-omics analysis revealed the mitochondrial-targeted drug combination to suppress the development of lung cancer.

Chaoqun Li, Yanfei Zhang, Qing Xia, Bingjie Hao, Yifan Hong, Liduo Yue, Tiansheng Zheng, Ming Li, Lihong Fan.

   PURPOSE: The incidence and mortality of lung cancer are continuously rising in recent years. Mitochondrial energy metabolism malfunction is found to be crucial in cancer proliferation and bioenergetic reprogramming, especially for lung cancer. In this study, we attempted to use mitochondrial-targeted drug therapy to change the energy metabolism pattern of cancer cells to inhibit the development of lung cancer, and investigated its mechanism of action and key targets through multi-omics studies.
METHODS: In this study, we established the in vivo tumor mouse mode, treated mice with multiple mitochondrial-targeted drug combinations and DDP, severally. Then, we investigated the differences between the 7-drug group with the control group and the DDP treatment group by transcriptomics, proteomics and metabolomics to find the therapeutic targets.
RESULTS: We found that mitochondria-targeting drug cocktail therapy, especially the 7-drug regimen, effectively improved mitochondrial metabolism, changed energy supply patterns in lung cancer cells, significantly increased NK cells in tumor tissues, and decreased tumor markers in plasma. Multi-omics analysis informed that the combination of 7-drug could up-regulate mitochondrial oxidative phosphorylation, ATP synthesis and autophagy related genes, and down-regulate proliferation and immune-related genes compared with the control group. By further mapping the protein interaction network, we identified a key target for 7-drug therapy to reverse tumor metabolic reprogramming and validated it in metabolomics.
CONCLUSIONS: Mitochondrial-targeted drug cocktail therapy can effectively inhibit the occurrence and development of tumors, through the reprogramming of energy metabolism and the increase in immune cells in tumor tissues. Thus, we provide a novel approach for the treatment of lung cancer and present evidence-based clues for the combined use of targeted mitochondrial drugs.

Keywords:  Cocktail therapy; Energy metabolism; Lung cancer; Mitochondria; Mitochondria targeting drug; Omics analysis

DOI:  https://doi.org/10.1007/s00432-023-05376-9
Biomed Pharmacother. 2023 Oct 04. pii: S0753-3322(23)01403-8. [Epub ahead of print]167 115605

Aminooxyacetic acid hemihydrochloride leads to decreased intracellular ATP levels and altered cell cycle of prostate cancer cells by suppressing energy metabolism.

Hailong Teng, Biao Yang, Yuangang Su, Junchun Chen, Lei Cui, Ran Sun, Jinmin Zhao, Qian Liu, An Qin.

  The second most common cancer among men is prostate cancer, which is also the fifth leading reason for male cancer deaths worldwide. Bone metastases are the main factor affecting the prognosis of prostate cancer. Consequently, antitumor and anti-prostate cancer-induced bone destruction medicines are urgently needed. We previously discovered that aminooxyacetic acid hemihydrochloride (AOAA) suppressed bone resorption and osteoclast growth by decreasing adenosine triphosphate (ATP) production and limiting oxidative phosphorylation (OXPHOS). Here, we evaluated the impacts of AOAA on prostate cancer RM-1 cells in vitro. It's found that AOAA significantly inhibited cell proliferation, migration, and invasiveness, decreased ATP levels, increased ROS, halted the cell cycle phase, and triggered apoptosis. AOAA also decreased mitochondrial membrane potential and the ability to uptake glucose, suggesting that the antitumor effects of AOAA were expressed through the inhibition of OXPHOS and glycolysis. Furthermore, we assessed the effects of AOAA in vivo using a prostate cancer-induced bone osteolysis mice model. AOAA also delayed tumor growth and bone destruction in vivo. On the whole, our findings imply that AOAA may potentially have therapeutic effects on prostate cancer and prostate cancer-induced osteolysis.

Keywords:  Aminooxyacetic acid hemihydrochloride; Apoptosis; Cell cycle; Energy metabolism; Prostate cancer; Reactive oxygen species

DOI:  https://doi.org/10.1016/j.biopha.2023.115605
Front Cell Dev Biol. 2023 ;11 1254313

Rewiring of mitochondrial metabolism in therapy-resistant cancers: permanent and plastic adaptations.

Katherine E Pendleton, Karen Wang, Gloria V Echeverria.

  Deregulation of tumor cell metabolism is widely recognized as a "hallmark of cancer." Many of the selective pressures encountered by tumor cells, such as exposure to anticancer therapies, navigation of the metastatic cascade, and communication with the tumor microenvironment, can elicit further rewiring of tumor cell metabolism. Furthermore, phenotypic plasticity has been recently appreciated as an emerging "hallmark of cancer." Mitochondria are dynamic organelles and central hubs of metabolism whose roles in cancers have been a major focus of numerous studies. Importantly, therapeutic approaches targeting mitochondria are being developed. Interestingly, both plastic (i.e., reversible) and permanent (i.e., stable) metabolic adaptations have been observed following exposure to anticancer therapeutics. Understanding the plastic or permanent nature of these mechanisms is of crucial importance for devising the initiation, duration, and sequential nature of metabolism-targeting therapies. In this review, we compare permanent and plastic mitochondrial mechanisms driving therapy resistance. We also discuss experimental models of therapy-induced metabolic adaptation, therapeutic implications for targeting permanent and plastic metabolic states, and clinical implications of metabolic adaptations. While the plasticity of metabolic adaptations can make effective therapeutic treatment challenging, understanding the mechanisms behind these plastic phenotypes may lead to promising clinical interventions that will ultimately lead to better overall care for cancer patients.

Keywords:  cancer; metabolism; oxphos (oxidative phosphorylation); plasticity; resistance

DOI:  https://doi.org/10.3389/fcell.2023.1254313
Cell Rep. 2023 Oct 03. pii: S2211-1247(23)01203-2. [Epub ahead of print]42(10): 113191

Spatiotemporal modeling of chemoresistance evolution in breast tumors uncovers dependencies on SLC38A7 and SLC46A1.

Yannick Audet-Delage, Catherine St-Louis, Lucía Minarrieta, Shawn McGuirk, Irwin Kurreal, Matthew G Annis, Arvind Singh Mer, Peter M Siegel, Julie St-Pierre.

  In solid tumors, drug concentrations decrease with distance from blood vessels. However, cellular adaptations accompanying the gradated exposure of cancer cells to drugs are largely unknown. Here, we modeled the spatiotemporal changes promoting chemotherapy resistance in breast cancer. Using pairwise cell competition assays at each step during the acquisition of chemoresistance, we reveal an important priming phase that renders cancer cells previously exposed to sublethal drug concentrations refractory to dose escalation. Therapy-resistant cells throughout the concentration gradient display higher expression of the solute carriers SLC38A7 and SLC46A1 and elevated intracellular concentrations of their associated metabolites. Reduced levels of SLC38A7 and SLC46A1 diminish the proliferative potential of cancer cells, and elevated expression of these SLCs in breast tumors from patients correlates with reduced survival. Our work provides mechanistic evidence to support dose-intensive treatment modalities for patients with solid tumors and reveals two members of the SLC family as potential actionable targets.

Keywords:  CP: Cancer; CP: Metabolism; anthracyclines; breast cancer; cancer metabolism; chemotherapy resistance; metabolomics; solute carriers

DOI:  https://doi.org/10.1016/j.celrep.2023.113191
Cancer Immunol Immunother. 2023 Oct 06.

Intensified NK cell therapy in combination with low-dose chemoradiotherapy against human colorectal cancer.

Huy Phuoc Quang Nguyen, Woo Kyun Bae, Myong Suk Park, Ik-Joo Chung, Taek-Keun Nam, Jae-Uk Jeong, Tung Nguyen Thanh Uong, Duck Cho, Sang-Ki Kim, Meesun Yoon.

  The therapeutic potential of adoptive natural Killer (NK) cells immunotherapy in combination with chemoradiotherapy, the main treatment modality for colorectal cancer (CRC), has not yet been explored. Here, we aimed to investigate the efficacy of NK cells to potentiate primary tumor control and improve survival outcomes, especially in combination with low-dose chemoradiotherapy. Ex vivo activated NK cells (> 90% purity) from healthy donors were obtained. NK cells were administered intravenously to the CRC-bearing mice and intensified in vivo in combination with low-dose 5-fluorouracil (0.5 mg/kg or 1 mg/Kg) and irradiated tumors with low doses (2 Gy or 4 Gy). Real-time NK cell cytotoxicity demonstrated a synergistic killing effect of a combination of low-dose chemoradiotherapy, mainly through NKp30 and NKG2D, showing a decrease in NK cell degranulation after blocking NKG2D and NKp30. In vivo tumor characteristics after combination treatment showed decreased CD112, CD155, MICA, and MICB expression. Under the combination strategy, 70% of the mice had free lung metastasis and 90% without secondary gross tumors, indicating suppressed distant metastasis to lung and axillary regions. This combination therapy resulted in significantly synergistic antitumor activity against primary solid tumors compared to chemoradiotherapy only. Furthermore, the intensified NK cell administration showed significantly better primary tumor control and survival outcomes than the non-intensified NK cell administration in a human colorectal HT-29 model treated with low-dose chemoradiotherapy. Optimized NK cell therapy combined with low-dose chemoradiotherapy can provide effective therapeutic potential for intractable cold human colorectal cancer.

Keywords:  Chemotherapy; Colorectal cancer; NK cell; Radiotherapy; Tumor microenvironment

DOI:  https://doi.org/10.1007/s00262-023-03545-w