bims-meluca Biomed News
on Metabolism of non-small cell lung carcinoma
Issue of 2022–05–15
twenty papers selected by
the Muñoz-Pinedo/Nadal (PReTT) lab, L’Institut d’Investigació Biomèdica de Bellvitge and Cristina Muñoz Pinedo, L’Institut d’Investigació Biomèdica de Bellvitge



  1. Clin Exp Immunol. 2022 May 13. 208(1): 60-71
      First-line immune checkpoint inhibitors (ICIs) have greatly ameliorated outcomes in non-small cell lung cancer (NSCLC). However, approximately a quarter of patients receiving ICIs demonstrate long-term clinical benefit, and the true responders have not been fully clarified by the existing biomarkers. To discover potential biomarkers treatment-related outcomes in plasma, mass spectrometry assay for the data-independent acquisition was analyzed plasma samples collected before the anti-PD-1 treatment. From July 2019 to January 2020, 15 patients with EGFR/ALK-negative NSCLC receiving first-line anti-programmed cell death protein 1 (PD-1) inhibitors were enrolled, and six healthy individuals have collected the plasma samples as control. We explored plasma proteome profiles and conducted stratified analyses by anti-PD-1 responders and non-responders. To validate the target proteins by ELISA, we recruited 22 additional independent patients and 15 healthy individuals from April 2021 to August 2021. By identifying biomarkers to predict better efficacy, we performed differential expression analysis in 12 responders and three non-responders. Compared with healthy individuals, hierarchical cluster analysis revealed plasma proteome profiles of NSCLC were markedly changed in 170 differentially expressed proteins. Furthermore, we discovered that SAA1, SAA2, S100A8, and S100A9 were noticeably increased among non-responders than responders, which may serve as predictive biomarkers with unfavorable responses. The validated results from all samples via ELISA have confirmed this observation. Identified a set of plasma-derived protein biomarkers (SAA1, SAA2, S100A8, and S100A9) that could potentially predict the efficacy in cohorts of patients with NSCLC treated with first-line anti-PD-1 inhibitors and deserves further prospective study.
    Keywords:  data-independent acquisition; immunotherapy; non-small cell lung cancer; proteomics; tumor biomarkers
    DOI:  https://doi.org/10.1093/cei/uxac021
  2. BMC Med. 2022 May 13. 20(1): 187
       BACKGROUND: Cytokines have been reported to alter the response to immune checkpoint inhibitors (ICIs) in patients with the tumor in accordance with their plasma concentrations. Here, we aimed to identify the key cytokines which influenced the responses and stimulated resistance to ICIs and tried to improve immunological response and develop novel clinical treatments in non-small cell lung cancer (NSCLC).
    METHODS: The promising predictive cytokines were analyzed via the multi-analyte flow assay. Next, we explored the correlation baseline level of plasma cytokines and clinical outcomes in 45 NSCLC patients treated with ICIs. The mechanism of the potential candidate cytokine in predicting response and inducing resistance to ICIs was then investigated.
    RESULTS: We found NSCLC with a low baseline concentration of IL-6 in plasma specimens or tumor tissues could derive more benefit from ICIs based on the patient cohort. Further analyses revealed that a favorable relationship between PD-L1 and IL-6 expression was seen in NSCLC specimens. Results in vitro showed that PD-L1 expression in the tumor was enhanced by IL-6 via the JAK1/Stat3 pathway, which induced immune evasion. Notably, an adverse correlation was found between IL-6 levels and CD8+ T cells. And a positive association between IL-6 levels and myeloid-derived suppressor cells, M2 macrophages and regulator T cells was also seen in tumor samples, which may result in an inferior response to ICIs. Results of murine models of NSCLC suggested that the dual blockade of IL-6 and PD-L1 attenuated tumor growth. Further analyses detected that the inhibitor of IL-6 stimulated the infiltration of CD8+ T cells and yielded the inflammatory phenotype.
    CONCLUSIONS: This study elucidated the role of baseline IL-6 levels in predicting the responses and promoting resistance to immunotherapy in patients with NSCLC. Our results indicated that the treatment targeting IL-6 may be beneficial for ICIs in NSCLC.
    Keywords:  Clinical response; IL-6; Immune checkpoint inhibitors; NSCLC; Resistance
    DOI:  https://doi.org/10.1186/s12916-022-02356-7
  3. Cell Commun Signal. 2022 May 09. 20(1): 62
       BACKGROUND: Recent studies have indicated that some members of the tripartite motif (TRIM) proteins function as important regulators for non-small cell lung cancer (NSCLC), However, the regulatory mechanism underpinning aberrant expression of TRIM in NSCLC remains unclear. Here we report that TRIM15 plays important roles in NSCLC progression through modulating Keap1-Nrf2 signaling pathway.
    METHODS: TRIM15 expression was evaluated by western blot analysis, tissue microarray-based immunohistochemistry analysis. The interactions between TRIM15 and Keap1 were analyzed by co-immunoprecipitation (Co-IP) and immunofluorescence co-localization assay. The correlation between TRIM15 and Keap1 was measured by Co-IP and ubiquitination analysis in vitro. Gain- and lost-of-function experiments were used to detect TRIM15 promotes proliferation and invasion of NSCLC cells both in vitro and vivo.
    RESULTS: Here, we revealed that TRIM15 was frequently upregulated in NSCLC samples and associated with poor prognosis. Functionally, TRIM15 knockdown resulted in decreased cancer cell proliferation and metastasis, whereas ectopic TRIM15 expression facilitated tumor cancer cell proliferation and metastasis in vitro and in vivo. Moreover, TRIM15 promoted cell proliferation and metastasis depends on its E3 ubiquitin ligase. Mechanistically, TRIM15 directly targeted Keap1 by ubiquitination and degradation, the principal regulator of Nrf2 degradation, leading to Nrf2 escaping from Keap1-mediated degradation, subsequently promoting antioxidant response and tumor progression.
    CONCLUSIONS: Therefore, our study characterizes the pivotal roles of TRIM15 promotes NSCLC progression via Nrf2 stability mediated by promoting Keap1 ubiquitination and degradation and could be a valuable prognostic biomarker and a potential therapeutic target in NSCLC. Video Abstract.
    Keywords:  Keap1; NSCLC; Nrf2; TRIM15; Ubiquitination and degradation
    DOI:  https://doi.org/10.1186/s12964-022-00875-7
  4. Nutr Cancer. 2022 May 11. 1-8
       PURPOSE: The aim of this study was to investigate the effect of the body fat mass ratio on survival and prognosis in advanced non-small-cell lung cancer patients.
    METHODS: The study includes 200 patients who were diagnosed with advanced non-small-cell lung cancer between 2014 and 2018 and whose body fat mass percentage and body mass index (BMI) were determined using the Tanita Body Composition Analyzer during admission.
    RESULTS: All patients had advanced incurable non-small-cell lung cancer (30% had locally advanced disease, 70% were stage IV). In the univariate and multivariate analyses, age, gender, histopathological type, smoking history, comorbidities, weight loss in the last six months and body mass index had no statistically significant effect on survival (p > 0.05). However, the performance status (p = 0.008), metastatic status (p = 0.003) and body fat mass ratio (p = 0.01) were found to have a significant effect on overall survival (OS): the median OS was 16.4 mo, in patients with the BFM ratio ≤ 22% and 29.2 mo, in those with > 22% (p = 0.01).
    CONCLUSION: In this study, it was found that the body fat mass ratio was an important prognostic factor in patients with advanced non-small-cell lung cancer.
    DOI:  https://doi.org/10.1080/01635581.2022.2074064
  5. Cancers (Basel). 2022 Apr 20. pii: 2063. [Epub ahead of print]14(9):
      There is an urgent need to identify new predictive biomarkers for treatment response to both platinum doublet chemotherapy (PT) and immune checkpoint blockade (ICB). Here, we evaluated whether treatment outcome could be affected by KRAS mutational status in patients with metastatic (Stage IV) non-small cell lung cancer (NSCLC). All consecutive patients molecularly assessed and diagnosed between 2016-2018 with Stage IV NSCLC in the region of West Sweden were included in this multi-center retrospective study. The primary study outcome was overall survival (OS). Out of 580 Stage IV NSCLC patients, 35.5% harbored an activating mutation in the KRAS gene (KRASMUT). Compared to KRAS wild-type (KRASWT), KRASMUT was a negative factor for OS (p = 0.014). On multivariate analysis, KRASMUT persisted as a negative factor for OS (HR 1.478, 95% CI 1.207-1.709, p < 0.001). When treated with first-line platinum doublet (n = 195), KRASMUT was a negative factor for survival (p = 0.018), with median OS of 9 months vs. KRASWT at 11 months. On multivariate analysis, KRASMUT persisted as a negative factor for OS (HR 1.564, 95% CI 1.124-2.177, p = 0.008). KRASMUT patients with high PD-L1 expression (PD-L1high) had better OS than PD-L1highKRASWT patients (p = 0.036). In response to first-line ICB, KRASMUT patients had a significantly (p = 0.006) better outcome than KRASWT patients, with a median OS of 23 vs. 6 months. On multivariable Cox analysis, KRASMUT status was an independent prognostic factor for better OS (HR 0.349, 95% CI 0.148-0.822, p = 0.016). kRAS mutations are associated with better response to treatment with immune checkpoint blockade and worse response to platinum doublet chemotherapy as well as shorter general OS in Stage IV NSCLC.
    Keywords:  KRAS; biomarker; chemotherapy; immunotherapy; lung cancer
    DOI:  https://doi.org/10.3390/cancers14092063
  6. FASEB J. 2022 May;36 Suppl 1
      Lung adenocarcinoma (LUAD) is the major histological subgroup of non-small cell lung cancer (NSCLC) with an extremely high mortality rate even when diagnosed at an early stage. LUAD makes up around 40% of lung cancer cases and can quickly metastasize to other areas of the body. Glycogen is the short-term storage of carbohydrates and can be easily mobilized in the body. Recently, other and our own data suggests glycogen can drive lung cancer progression, however, the mechanism of which glycogen accumulates in LUAD is poorly understood and potentially arises from a number of genetic and environmental factors. Although specific genetic variants driving aberrant LUAD glycogen accumulation have been identified, several studies have shown that caloric excess as well as specific nutrients in the diet can promote tumor growth. Further, LUAD patients with type II diabetes or high BMI have significantly poorer survival. Therefore, we hypothesized that a diet rich in fats and carbohydrates that mimics a Western diet increases glycogen accumulation in lung tissue and potentially drives LUAD tumor progression. To test our hypothesis that a Western diet alters glycogen metabolism independent of genetic drivers and tumor formation, we administered via oral gavage a Western diet comprised of corn oil and high fructose corn syrup (150 μl corn oil: 25% high-fructose corn syrup solution) to wild-type C57BL/6 mice at acute (1-, 3-, and 6-hours) and chronic (two weeks) time points. At the end of each time point, mice were sacrificed and lung tissue was harvested. Using a combination of gas chromatography mass spectrometry (GCMS) and matrix-assisted laser desorption ionization mass spectrometry imaging, we quantified central carbon metabolites and glycogen content in lung tissue resected from these mice. Mice administered H2O were used as a control. We found that the mice administered a Western diet for two weeks exhibited elevated levels of glycogen as well as metabolite pools within glycolysis and the TCA cycle in lung tissue compared to the control group. However, our acute studies revealed no change in lung glycogen up to 6-hours after receiving the Western diet. Overall, these data highlight a link between diet and glycogen metabolism in the lung and suggest that intake of different dietary nutrients may play a role in glycogen accumulation in LUAD tumors and disease progression. Our results demonstrate that a high fat/high carbohydrate diet in wild-type mice increases central carbon metabolism and glycogen accumulation in lung tissue after chronic exposure. Moving forward, we will examine the contribution of diet to glycogen metabolism in a mouse of LUAD and assess the effect of increased glycogen accumulation on tumor growth in vivo.
    DOI:  https://doi.org/10.1096/fasebj.2022.36.S1.R2115
  7. Oncol Lett. 2022 Jun;23(6): 184
      Aerobic glycolysis plays a key role in cancer cell metabolism and contributes to tumorigenesis, including that of non-small cell lung cancer (NSCLC). Tanshinone IIA (Tan IIA), an active compound of Salvia miltiorrhiza, exhibits antitumor properties. Multiple mechanisms are involved in the antitumor action of Tan IIA in lung cancer, such as inhibiting cell growth, promoting cell apoptosis and influencing cellular metabolism. However, the effects of Tan IIA on NSCLC cells and its mechanisms of action remain unclear. The present study shows Tan IIA dose-dependently attenuated the growth of NSCLC cells and in vitro in a dose-dependent manner. Moreover, Tan IIA markedly decreased the ATP level, glucose uptake and lactate production in the NSCLC cells in vitro. Tan IIA also inhibited tumor growth in a xenograft model in vivo. Mechanically, Tan IIA treatment decreased sine oculis homeobox homolog 1 (SIX1) mRNA and protein levels, thus leading to the downregulation of pyruvate kinase isozyme M2, hexokinase 2 and lactate dehydrogenase A (LDHA) expression in A549 cells. SIX1 knockdown with small interfering-RNA inhibited glycolysis in NSCLC cells, suggesting that SIX1 plays a role in the antitumor effect of Tan IIA on NSCLC cells. More importantly, it was demonstrated that SIX1 expression was stimulated in patients with NSCLC and was positively correlated with the LDH serum level. Finally, SIX1 low expression levels predicted the poor prognosis of patients with NSCLC. In conclusion, the present study showed that Tan IIA functioned as an anti-glycolysis agent in NSCLC cells by downregulating SIX1 expression and inhibiting cell proliferation.
    Keywords:  SIX1; Tan IIA; aerobic glycolysis; non-small cell lung cancer; proliferation
    DOI:  https://doi.org/10.3892/ol.2022.13304
  8. Oxid Med Cell Longev. 2022 ;2022 2590198
      The tryptophan residue has a large hydrophobic surface that plays a unique role in the folded protein conformation and functions. Tryptophan modifications are presumably to be readily detected in proteins due to the vulnerability of the indole structure to electrophilic attacks. In this study, we report a systematic identification of sequence variations at tryptophan, termed tryptophan variants, from the proteome of patients with nonsmall cell lung cancer (NSCLC). Using shotgun proteomics and a modified open search algorithm, 25 tryptophan variants on 2481 sites in over 858 proteins were identified. Among these, 6 tryptophan variants are previously identified, 15 are newly annotated, and 4 are still unknown, most of which are involved in the cascade of oxidation in the blood microparticle. Remarkably, Trp313 of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was up-oxidized whereas Trp16 and Trp38 of hemoglobin (HBB) were down-oxidized in NCSLC tissues. The results were further supported by an independent cohort of 103 lung adenocarcinoma samples, reflecting a negative feedback and potential detoxification mechanism against tumor glycolysis and hypoxia. Overall, the study reports a quick approach to explore tryptophan variants at the proteomic scale. Our findings highlight the predominant role of tryptophan oxidation in regulating the redox balance of cancer cells and its potential role as prognostic biomarker for patients with NSCLC.
    DOI:  https://doi.org/10.1155/2022/2590198
  9. Life Sci. 2022 May 08. pii: S0024-3205(22)00321-6. [Epub ahead of print] 120621
       AIMS: Lung cancer is the leading cause of cancer-related death. Unfortunately, targeted-therapies have been unsuccessful for most patients with lung adenocarcinoma (LUAD). Thus, new early biomarkers and treatment options are a pressing need. Fatty acid binding protein 5 (FABP5) has been associated with various types of cancers. Its contribution to LUAD onset, progression and metabolic reprogramming is, however, not fully understood. In this study we assessed the importance of FABP5 in LUAD and its role in cancer lipid metabolism.
    MAIN METHODS: By radioactive labeling and metabolite quantification, we studied the function of FABP5 in fatty acid metabolism using genetic/pharmacologic inhibition and overexpression models in LUAD cell lines. Flow cytometry, heterologous transplantation and bioinformatic analysis were used, in combination with other methodologies, to assess the importance of FABP5 for cellular proliferation in vitro and in vivo and in patient survival.
    KEY FINDINGS: We show that high expression of FABP5 is associated with poor prognosis in patients with LUAD. FABP5 regulates lipid metabolism, diverting fatty acids towards complex lipid synthesis, whereas it does not affect their catabolism in vitro. Moreover, FABP5 is required for de novo fatty acid synthesis and regulates the expression of enzymes involved in the pathway (including FASN and SCD1). Consistently with the changes in lipid metabolism, FABP5 is required for cell cycle progression, migration and in vivo tumor growth.
    SIGNIFICANCE: Our results suggest that FABP5 is a regulatory hub of lipid metabolism and tumor progression in LUAD, placing it as a new putative therapeutic target for this disease.
    Keywords:  FABP5; Fatty acid biosynthesis; Lipogenesis; Lung cancer; Metabolism
    DOI:  https://doi.org/10.1016/j.lfs.2022.120621
  10. FASEB J. 2022 May;36 Suppl 1
      Metabolism reprogramming and excessive glycolysis are associated with rapid cancer cell growth, motility, and resistance to drug treatment. Therefore, identifying candidates for regulating the ratio of glycolysis and gluconeogenesis is a priority. Especially the reverse effect of Aldolase/Fructose-1,6-Bisphosphatase. Fructose-1, 6-bisphosphate (F-1,6-BP), is a vital intermediate involvement in glucose metabolism. When F-1,6-BP is converted into the 3-carbon product by Aldolase, the glycolysis will proceed sequentially, while FBP1 can reverse the equation. Based on established immunoprecipitation and proteomic analyses revealed several candidates, including the anterior gradient 2 (AGR2), ubiquitin ligase gene cullin 4B (CUL4B), and telomeric repeat binding factor 1 (TERF1) that significantly modulates the ratio between ALDOA and FBP1. Our study demonstrated the inversed correlation of ALDOA and FBP1 associated with lung adenocarcinoma and hepatocellular carcinoma progression. Molecular simulated analysis and clinical correlation indicated that CUL4B is the novel factor in switching ALDOA and FBP1. Moreover, we evaluate the CUL4B-relative glucose metabolism events to describe how CUL4B participates in the switch of ALDOA and FBP1. Based on these relationships, we claimed that CUL4B could trigger glucose metabolism reprogramming and be used as a therapeutic target for further research.
    DOI:  https://doi.org/10.1096/fasebj.2022.36.S1.R3282
  11. FASEB J. 2022 May;36 Suppl 1
      Lung cancer is one of the most prevalent cancers worldwide, accounting for nearly two million new cases each year. Lung adenocarcinoma (LUAD) is the most common subtype of lung cancer and is the leading cause of cancer related mortalities worldwide. LUAD has an extremely poor five-year survival rate, which is likely due to cases being diagnosed at advanced stages and thus being more difficult to treat the metastatic disease. Identifying the molecular processes that contribute to disease progression is a critical goal in lung cancer research that could lead to predictive biomarkers and novel therapies. Glycogen is the primary form of carbohydrate storage in mammalian cells, and its degradation product, Glucose 6-Phosphate (G6P) feeds directly into glycolysis, making glycogen intimately connected to central carbon metabolism. Recently, aberrant glycogen accumulation in lung tumors has been reported to promote lung cancer progression, driven by largely unknown mechanisms. We recently generated a transgenic KRASG12D /p53-/- LUAD mouse model that also lacks the glycogen phosphatase laforin (LKO). This model develops increased glycogen accumulation in the lungs and displays accelerated tumor growth compared to the KRASG12D /p53-/- control (WT), as measured by Ki67 staining. To identify potential biological processes perturbed by LUAD-glycogen, we performed RNA-sequencing on lung tumors resected from WT and LKO mice. We identified over 300 genes that are differentially expressed between WT and LKO lung tumors. Specifically, gene set enrichment analysis (GSEA) revealed genes that were up-regulated in LKO lung tumors were involved in extracellular matrix remodeling, cell-to-cell communication, and cell proliferation. All of these processes are implicated in tumor growth, which supports our initial observation that LKO mice have increased tumor burden and disease progression. Our findings ultimately highlight several potential mechanisms by which LUAD-glycogen promotes LUAD tumor progression, which can be further investigated to identify novel biomarkers and therapeutic targets.
    DOI:  https://doi.org/10.1096/fasebj.2022.36.S1.R3959
  12. Stem Cell Res Ther. 2022 May 12. 13(1): 197
       BACKGROUND: Chemoresistance often causes the failure of treatment and death of patients with advanced non-small-cell lung cancer. However, there is still no resistance genes signature and available enriched signaling derived from a comprehensive RNA-Seq data analysis of lung cancer patients that could act as a therapeutic target to re-sensitize the acquired resistant cancer cells to chemo-drugs. Hence, in this study, we aimed to identify the resistance signature for clinical lung cancer patients and explore the regulatory mechanism.
    METHOD: Analysis of RNA-Seq data from clinical lung cancer patients was conducted in R studio to identify the resistance signature. The resistance signature was validated by survival time of lung cancer patients and qPCR in chemo-resistant cells. Cytokine application, small-interfering RNA and pharmacological inhibition approaches were applied to characterize the function and molecular mechanism of EREG and downstream signaling in chemoresistance regulation via stemness.
    RESULTS: The RTK and vitamin D signaling were enriched among resistance genes, where 6 genes were validated as resistance signature and associated with poor survival in patients. EREG/ERK signaling was activated by chemo-drugs in NSCLC cells. EREG protein promoted the NSCLC resistance to chemo-drugs by increasing stemness genes expression. Additionally, inhibition of EREG/ErbB had downregulated ERK signaling, resulting in decreased expression of stemness-associated genes and subsequently re-sensitized the resistant NSCLC cells and spheres to chemo-drugs.
    CONCLUSIONS: These findings revealed 6 resistance genes signature and proved that EREG/ErbB regulated the stemness to maintain chemoresistance of NSCLC via ERK signaling. Therefore, targeting EREG/ErbB might significantly and effectively resolve the chemoresistance issue.
    Keywords:  Chemoresistance; EREG; ERK signaling; Non-small cell lung cancer; Receptor tyrosine kinase; Stemness
    DOI:  https://doi.org/10.1186/s13287-022-02859-3
  13. Am J Cancer Res. 2022 ;12(4): 1535-1555
      Multi-drug resistance remains a critical issue in cancer treatment that hinders the effective use of chemotherapeutic drugs. The active components of traditional Chinese medicine have been applied as adjuvants to accentuate the anticancer properties of conventional drugs such as cisplatin. However, their application requires further validation and optimization. This study explored the anticancer activity of β-elemene, a natural component of traditional Chinese medical formulations. The effect of β-elemene on the anticancer properties of cisplatin was evaluated in A549 and NCI-H1650 lung cancer cells. Cell apoptosis, stem-like properties, glucose metabolism, multi-drug resistance, and PI3K/AKT/mTOR activation were assessed via flow cytometry, tumorsphere formation, and western blotting. The target genes of β-elemene were predicted using bioinformatics tools and validated in both cell lines. A xenograft model of lung cancer was established in nude mice to evaluate the combined effects of β-elemene and cisplatin in vivo. We found that β-elemene acted synergistically with cisplatin against non-small cell lung cancer cells by promoting apoptosis and impairing glucose metabolism, multi-drug resistance, and stemness maintenance. These effects were mediated by the inhibition of PI3K/AKT/mTOR activation. Bioinformatics analysis revealed that RB1 and TP53 are common target genes associated with lung cancer and β-elemene. The anti-tumorigenic properties of β-elemene were confirmed in vivo, wherein β-elemene, along with cisplatin, significantly suppressed tumor growth in a mouse xenograft model of non-small cell lung cancer. As such, β-elemene acted as an inhibitor of PI3K/AKT/mTOR signaling and enhanced the anticancer effect of cisplatin by targeting tumor metabolism, chemoresistance, and stem-like behavior. Thus, β-elemene is an effective anticancer adjuvant agent with potential clinical applications.
    Keywords:  Lung cancer; Warburg effect; apoptosis; cancer stem cell; multi-drug resistance; β-elemene
  14. FASEB J. 2022 May;36 Suppl 1
      Escaping of immune surveillance is essential for tumor metastasis. Recently, intratumoral metal ionic balance plays an important role in controlling cancer immune checkpoint protein expression and tumor immune escape. Copper (Cu) promotes the expression of PD-L1. In contrast to Cu, heme biosynthesis disturbs PD-L1 expression. Heme protein is a kind of metalloproteins with ferrous iron (Fe2+ )-porphyrin core. Here, we found CHST11 served as a pro-metastatic gene in non-small cell lung cancer (NSCLC). We further identified the expression of ceruloplasmin (CP) was regulated by CHST11. CP is not only the major blood Cu carrier protein but also a Cu-dependent ferroxidase in iron metabolism. It may hint a crosstalk mechanism between lung cancer metastasis and tumor immune surveillance through the CHST11-CP axis. CP was also associated with poor prognosis in patients with NSCLC. Both ectopic CP expression and high intracellular ionic stimulations, like ferric iron and copper, promoted NSCLC progression, such as in vitro migration and invasion, as well as in vivo tumorigenesis abilities in Nod-SCID mice. The ionic chelator treatments repressed in vitro metastatic potential and, interestingly, regulated the gene expressions of immune checkpoints in NSCLC cells. By using microarray analysis, we found CHST11 controlled CP expression through interferon-gamma (IFNγ), Janus kinase (JAK), and signal transducer and activator of transcription (STAT) pathways. Treatment with ruxolitinib, a JAK inhibitor, suppressed the expressions of CP and immune checkpoint proteins. In conclusion, our findings prove that the CHST11-CP axis plays an important role in both NSCLC immune escape and metastasis through IFNγ-JAK-STAT pathway. The combination of an iron chelator and ruxolitinib may provide a potential translation application that protects patients with NSCLC from cancer immune escape and metastasis.
    DOI:  https://doi.org/10.1096/fasebj.2022.36.S1.R5894
  15. FASEB J. 2022 May;36 Suppl 1
      Angiogenesis refers to the development of new blood vessels from preexisting blood vessels. The angiogenic pathway is essential for the growth and progression of almost every kind of human cancer. Nicotine, the addictive component of tobacco smoke is known to accelerate the growth of human lung tumors by stimulating tumor angiogenesis. The pro-angiogenic effects of nicotine are mediated by nicotinic acetylcholine receptors (nAChRs). The endogenous ligand for nicotine is the neurotransmitter acetylcholine (ACh). Published reports show that ACh acts as growth factor for human lung cancers. Almost all lung tissues (including fetal lung) express signaling proteins involved in the synthesis, transport of the ACh-signaling pathway. These include choline acetyl transferase enzyme (ChAT), vesicular acetylcholine transporter (VAChT), acetylcholine esterase (AChE) and choline transporter (ChT). The primary objective of our studies was to determine if disruption of ChAT could inhibit angiogenesis and suppress the growth of human LACs. We used a small molecule inhibitor of ChAT namely BW813U for our studies. BW813U robustly suppressed angiogenesis in Matrigel assays and chicken chorioallantoic membrane (CAM) assays. The administration of BW813U potenetly decreased the growth rate of H838 tumors xenotransplanted in SCID mice. Immunohistochemical staining experiments revealed that the anti-tumor activity of BW813U was correlated with decrease of CD31 angiogenic biomarker in H813 tumor sections. The anti-angiogenic activity of BW813U was mediated by the alpha7-nAChR pathway and involved the nitric oxide pathway. Taken together, our studies show that ChAT antagonists like BW813U may have applications in the treatment of LAC.
    DOI:  https://doi.org/10.1096/fasebj.2022.36.S1.L6388
  16. Am J Med Sci. 2022 May 04. pii: S0002-9629(22)00208-7. [Epub ahead of print]
      Glucocorticoids have powerful anti-inflammatory and immunomodulatory effects, but chronic use of these drugs can cause hyperglycemia, type 2 diabetes mellitus, hepatic steatosis, obesity, and other complications due to their metabolic actions. Metformin is a widely used drug for the treatment of type 2 diabetes mellitus with a known ability to lower blood glucose levels. This review focuses on metformin's actions on glucose metabolism and its potential use as a drug to limit the metabolic side effects of glucocorticoid treatment. Available data suggest that metformin inhibits complex I of the mitochondrial electron transport chain, crucial gluconeogenic enzymes, and fatty acid synthesis that leads to a significant improvement in glucose tolerance and maintenance of insulin sensitivity during glucocorticoid treatment. Three small randomized control trials have demonstrated that metformin can limit changes in glucose metabolism during treatment with prednisone. These studies reveal a promising potential for metformin use as a therapeutic agent to reduce glucocorticoid-induced hyperglycemia and improve patient outcomes.
    Keywords:  Metformin; glucocorticoids; glucose metabolism; hyperglycemia; new applications; treatment
    DOI:  https://doi.org/10.1016/j.amjms.2022.04.027
  17. Metabolomics. 2022 May 14. 18(5): 31
       INTRODUCTION: Metabolomics has emerged as a powerful method to provide insight into cancer progression, including separating patients into low- and high-risk groups for overall (OS) and progression-free survival (PFS). However, survival prediction based mainly on metabolites obtained from biofluids remains elusive.
    OBJECTIVES: This proof-of-concept study evaluates metabolites as biomarkers obtained directly from tumor core biopsies along with covariates age, sex, pathological stage at diagnosis (I/II vs. III/VI), histological subtype, and treatment vs. no treatment to risk stratify lung cancer patients in terms of OS and PFS.
    METHODS: Tumor core biopsy samples obtained during routine lung cancer patient care at the University of Louisville Hospital and Norton Hospital were evaluated with high-resolution 2DLC-MS/MS, and the data were analyzed by Kaplan-Meier survival analysis and Cox proportional hazards regression. A linear equation was developed to stratify patients into low and high risk groups based on log-transformed intensities of key metabolites. Sparse partial least squares discriminant analysis (SPLS-DA) was performed to predict OS and PFS events.
    RESULTS: Univariable Cox proportional hazards regression model coefficients divided by the standard errors were used as weight coefficients multiplied by log-transformed metabolite intensity, then summed to generate a risk score for each patient. Risk scores based on 10 metabolites for OS and 5 metabolites for PFS were significant predictors of survival. Risk scores were validated with SPLS-DA classification model (AUROC 0.868 for OS and AUROC 0.755 for PFS, when combined with covariates).
    CONCLUSION: Metabolomic analysis of lung tumor core biopsies has the potential to differentiate patients into low- and high-risk groups based on OS and PFS events and probability.
    Keywords:  Lung cancer; Metabolomics; Overall survival; Progression free survival; Risk score calculator; Tumor core biopsy
    DOI:  https://doi.org/10.1007/s11306-022-01891-x
  18. FASEB J. 2022 May;36 Suppl 1
      Lung adenocarcinoma is the most common type of lung cancer and has a strong association with tobacco smoking. Smoking leads to the accumulation of reactive oxygen species (ROS) and oxidative stress damages to macromolecules, leading to abnormal cell growth & proliferation, transformation, changes in cell metabolism and a variety of other physiological functions. Accumulation of ROS only leads to increased expression of cellular antioxidants that contribute to tumorigenesis and cancer progression. Sulfiredoxin (Srx) is the only enzyme that reduces over-oxidized forms of typical 2-Cys Peroxiredoxins (Prxs). Compared to other Prx family members, Srx preferentially interacts with and has a higher binding affinity to Prx4. Our previously work demonstrated that the Srx-Prx4 axis enhances the RAS/RAF-MEK-ERK signaling cascade to promote lung cancer cell proliferation and metastasis. However, the role of the Srx-Prx axis in de novo lung tumorigenesis has not been explored. In this study, Prx4-/- mice and Srx-/-Prx4-/- mice were generated and mouse lung carcinogenesis was induced by a well-established urethane protocol. Briefly, wildtype and knockout mice at 8-week of age were injected intraperitoneally with 1g/kg of urethane once per week for three successive weeks. Ten weeks after the administration of urethane, all mice were euthanized and examined for tumors. Our results indicate that both knockout mice have significantly reduced rates on tumor incidence, multiplicity, and size. Compared with tumors from wildtype mice, the rate of cell proliferation in tumors from either knockout mice is significantly decreased. Macrophages are the most abundant immune cells identified in the tumor microenvironment of solid tumors and their presence correlates with reduced survival in most cancers. In addition to all other markers, we also compared the presence of two distinct states of polarized macrophages including the classically activated macrophage (M1) and the alternatively activated macrophage subsets (M2) in urethane-induced tumors using specific surface marker staining. We found that there are significantly increased numbers of M1 cells in tumors of either Prx4-/- or Srx-/-Prx4-/- mice compared with those of wildtype mice. In contrast, there are significantly few numbers of M2 cells in tumors from either knockout mice. It has been shown that M1 macrophages have immune-stimulatory properties through the expression of a series of pro-inflammatory cytokines, chemokines, and effector molecules, whereas M2 cells express a wide array of anti-inflammatory molecules. Therefore, the difference in tumor-associated macrophages caused by the loss of Prx4 or Srx and Prx4 may contribute to the reduced lung tumorigenesis observed in the mouse model. In addition, bioinformatic analysis shows that Prx4 is overexpressed in the majority of cancers, and the increased expression of Srx and Prx4 both show a strong correlation with poor prognosis in lung adenocarcinoma patients.
    DOI:  https://doi.org/10.1096/fasebj.2022.36.S1.R5242
  19. Oncogenesis. 2022 May 09. 11(1): 24
      Lung cancer is the leading cause of cancer-related death worldwide despite the success of therapies targeting oncogenic drivers and immune-checkpoint inhibitors. Although metabolic enzymes offer additional targets for therapy, the precise metabolic proteome of lung adenocarcinomas is unknown, hampering its clinical translation. Herein, we used Reverse Phase Protein Arrays to quantify the changes in enzymes of glycolysis, oxidation of pyruvate, fatty acid metabolism, oxidative phosphorylation, antioxidant response and protein oxidative damage in 128 tumors and paired non-tumor adjacent tissue of lung adenocarcinomas to profile the proteome of metabolism. Steady-state levels of mitochondrial proteins of fatty acid oxidation, oxidative phosphorylation and of the antioxidant response are independent predictors of survival and/or of disease recurrence in lung adenocarcinoma patients. Next, we addressed the mechanisms by which the overexpression of ATPase Inhibitory Factor 1, the physiological inhibitor of oxidative phosphorylation, which is an independent predictor of disease recurrence, prevents metastatic disease. We highlight that IF1 overexpression promotes a more vulnerable and less invasive phenotype in lung adenocarcinoma cells. Finally, and as proof of concept, the therapeutic potential of targeting fatty acid assimilation or oxidation in combination with an inhibitor of oxidative phosphorylation was studied in mice bearing lung adenocarcinomas. The results revealed that this therapeutic approach significantly extended the lifespan and provided better welfare to mice than cisplatin treatments, supporting mitochondrial activities as targets of therapy in lung adenocarcinoma patients.
    DOI:  https://doi.org/10.1038/s41389-022-00400-y
  20. FASEB J. 2022 May;36 Suppl 1
      Although the overall prognosis for patients with non-small cell lung cancer (NSCLC) has improved over the past several decades, there are still survival differences that are not accurately defined by clinicopathological factors. Thus, there is an unmet clinical need to develop novel approaches to enhance prognostic accuracy for the assessment of these patients. Prior studies have established that keratin 17 (K17) is a negative prognostic biomarker in a wide range of cancer types, including pulmonary adenocarcinoma (LUAD). Although keratin expression profiles have been widely utilized as markers to distinguish between metastatic versus primary squamous cell carcinomas (SCC), K17 has not been previously investigated for its potential accuracy as a prognostic biomarker in lung squamous cell carcinoma (LSCC). Here, we set out to determine if K17 is a prognostic biomarker that drives poor survival in both LSCC and LUAD. Data mining was performed to analyze K17 mRNA in independent cohorts of LSCC (n=266) and LUAD (n=271) and immunohistochemistry (IHC) for K17 was performed in separate cohorts of LSCC (n=84) and LUAD (n=107). Kaplan-Meier and Cox proportional-hazard regression models were used to determine overall survival differences between low vs. high K17 expressing cases. High K17 mRNA correlated with decreased overall survival in both LSCC (HR: 1.789, p=0.0646) and LUAD (HR: 1.894, p=0.0142). Similarly, by IHC, high K17 correlated with decreased overall survival in LSCC (HR: 1.894, p=0.0375) and LUAD (HR: 2.105, p=0.0257). Thus, K17, defined either by mRNA expression or immunohistochemical staining, is a negative prognostic biomarker for non-small cell lung cancer.
    DOI:  https://doi.org/10.1096/fasebj.2022.36.S1.R2935