bims-meluca 2023-10-08 papers

on Metabolism of non-small cell lung carcinoma

Issue of 2023–10–08
seven papers selected by
the Muñoz-Pinedo/Nadal (PReTT) lab, L’Institut d’Investigació Biomèdica de Bellvitge

Circulating Growth Differentiation Factor 15 as a Biomarker in Patients with Unresected Locally Advanced Non-Small Cell Lung Cancer Treated with Chemo-Radiotherapy with or without Metformin.
High endothelial venules predict response to PD-1 inhibitors combined with anti-angiogenesis therapy in NSCLC.
Glucose restriction induces AMPK-SIRT1-mediated circadian clock gene Per expression and delays NSCLC progression.
Transcriptome-level Discovery of Survival-Associated Biomarkers and Therapy Targets in Non-Small-Cell Lung Cancer.
Prognostic Significance of Serum Lipids in Patients with Non-Small Cell Lung Cancer Treated with Radiotherapy: A Multicenter Prospective Study.
Prognostic Value of Hypoxia Gene Expression Signature in Early-Stage Non-Small Cell Lung Cancer Patients Treated with Stereotactic Body Radiation Therapy.
Tumor mutation burden in the prognosis and response of lung cancer patients to immune-checkpoint inhibition therapies.

Int J Radiat Oncol Biol Phys. 2023 Oct 01. pii: S0360-3016(23)05112-X. [Epub ahead of print]117(2S): e14-e15

Circulating Growth Differentiation Factor 15 as a Biomarker in Patients with Unresected Locally Advanced Non-Small Cell Lung Cancer Treated with Chemo-Radiotherapy with or without Metformin.

F Di Pastena, G Pond, E Ahmadi, E E Tsakiridis, G R Steinberg, T Tsakiridis.

PURPOSE/OBJECTIVE(S): Growth differentiation factor 15 (GDF15) is a member of the TGFβ family suggested to have prognostic value in cancer. Plasma GDF15 levels are elevated in humans during cellular stress, chronic diseases, and cancer and in response to cytotoxic (chemotherapy and radiation) or metabolic (metformin) therapies. Currently, the effect of combined metformin and cytotoxic treatment on circulating GDF15 is not known, and it is unclear whether GDF15 can serve as a biomarker in patients receiving such therapy. To examine this question, we analyzed circulating GDF15 levels in stage IIIA or IIIB NSCLC patients that participated in the randomized phase II clinical trial OCOG-ALMERA (NCT02115464).
MATERIALS/METHODS: In OCOG-ALMERA patients were randomized to treatment with concurrent platinum-based chemotherapy and chest radiotherapy (60-66 Gy), with or without consolidation chemotherapy (CRT) or the same treatment plus metformin (2000mg/d). Dosimetric parameters and survival outcomes were collected by the trial. EDTA Plasma was collected from patients at baseline, 2 weeks into CRT treatment, at completion of CRT and 6 months after treatment initiation and assayed for GDF15 using an ELISA method. Statistical analyses were conducted to explore potential associations between plasma GDF15 with survival and dosimetric parameters.
RESULTS: Fifteen patients from the metformin arm and 18 from the control arms provided EDTA plasma for this analysis. Average baseline GDF15 levels for all participants were elevated compared to those reported for healthy individuals. Baseline plasma GDF15 was statistically prognostic for RFS and OS (hazard ratio = 1.19, 95% CI = 1.06 to 1.34, p = 0.005). GDF15 levels increased during CRT treatment reaching the highest value at the end of cytotoxic therapy. Addition of metformin to CRT treatment was associated with further increase in circulating GDF15 (p<0.001). Plasma GDF15 levels at 2 weeks of CRT were positively correlated with size of the radiotherapy clinical target volume and volumes of esophagus and heart receiving high dose radiotherapy.
CONCLUSION: This work suggests that GDF15 may be a promising prognostic marker to predict response to standard chemoradiation therapy in NSCLC. Future studies should aim to validate these results in larger datasets and examine whether GDF15 may have value as early biomarker of radiation toxicity in lung cancer patients.

DOI: https://doi.org/10.1016/j.ijrobp.2023.06.678
Sci Rep. 2023 09 30. 13(1): 16468

High endothelial venules predict response to PD-1 inhibitors combined with anti-angiogenesis therapy in NSCLC.

Dafu Ye, Yao Jin, Yiming Weng, Xue Cui, Jinsong Wang, Min Peng, Qibin Song.

Tumor-associated high endothelial venules (TA-HEVs) mediate lymphocyte entry into tumors. Therefore, combined anti-angiogenesis therapy and programmed death-1 (PD-1) inhibitors might stimulate tumor immunity. This study will explore the TA-HEVs and real-world data of the combination therapy in non-small cell lung cancer (NSCLC). Firstly, we found a certain relationship between HEVs and immune effector cells by multiple immunofluorescence staining. We then analyzed the efficacy of immunotherapy combined with anti-angiogenesis therapy in advanced NSCLC patients by collecting real-world clinical data. Finally, we explored the predictive value of HEVs in combination therapy by analyzing pre-treatment pathological slides of patients with multiple immunofluorescence and RNA sequencing. Immunofluorescence staining of high endothelial venules (PNAd+) reveals that the frequency of HEVs is positively correlated with tumor-infiltrating stem-like CD8+ T cells (TCF-1+PD-1+) in the TME of advanced NSCLC patients (P = 0.0221). We retrospectively analyzed the efficacy of 96 patients with advanced NSCLC who received PD-1 inhibitors combined with anti-angiogenesis therapy in the real-world. The median PFS of patients combined with anti-angiogenesis therapy was longer than that of patients without anti-angiogenesis therapy (9.7 vs 8.6 months, P = 0.041). Multiple immunofluorescence staining of tumor biopsies before treatment from 14 patients with advanced NSCLC reveals that PNAd+ is predictive of better response and survival upon PD-1 inhibitors combined with anti-angiogenesis therapy (P = 0.0274). In addition, we collected peripheral blood from an effective group of patients for RNA sequencing and found that immune cells activation-related gene expression scores were higher. Combined anti-angiogenic and anti-PD-1 therapy stimulates tumor immunity through TA-HEVs formation. TA-HEVs not only mediate immune cell entry into tumors, but also are associated with the efficacy of PD-1 inhibitors and anti-angiogenesis therapy in NSCLC.

DOI: https://doi.org/10.1038/s41598-023-43122-w
Cancer Lett. 2023 Sep 29. pii: S0304-3835(23)00375-0. [Epub ahead of print] 216424

Glucose restriction induces AMPK-SIRT1-mediated circadian clock gene Per expression and delays NSCLC progression.

Bohan Li, Qianfeng Chen, Yucong Feng, Tao Wei, Yuxia Zhong, Yuandie Zhang, Qing Feng.

  The rhythmic expression of the circadian clock is intimately linked to the health status of the body. Disturbed circadian clock rhythms might lead to a wide range of metabolic diseases and even cancers. Our previous study showed that glucose restriction was able to inhibit non-small cell lung cancer (NSCLC). In the current study, we found that glucose restriction enhanced apoptosis and cell growth delay in NSCLC cells. In addition, we used GEPIA database analysis to derive different effects of each circadian clock gene on lung cancer tissue. Among these circadian clock genes, Per (Period) is lowly expressed in cancer tissues and highly expressed in normal tissues. Moreover, the higher expression of Per in cancer patients has a better prognostic significance. Furthermore, we revealed that glucose restriction induced the expression of the circadian clock gene Per in NSCLC cells by upregulating SIRT1 (Sirtuin1) via activation of the energy response factor AMPK (AMP-activated protein kinase). Changes in Per expression following upregulation or downregulation of AMPK were consistent with AMPK expression. Additionally, a low-carbohydrate ketogenic diet significantly delayed tumor progression in a xenograft tumor model of severe combined immunodeficiency (SCID) mice. Meanwhile, the ketogenic diet increased the expression of AMPK, SIRT1 and Per in vivo. Besides, the ketogenic diet was found to restore the normal rhythmic level of Per by Zeitgeber Time (ZT) experiments. Taken these together, these results indicated a novel mechanism that glucose restriction induces AMPK-SIRT1 mediated circadian clock gene Per expression and delays NSCLC progression, which provided more evidence for glucose restriction as an adjuvant clinical therapeutic strategy in NSCLC.

Keywords:  AMPK; Glucose restriction; NSCLC; Per; SIRT1

DOI:  https://doi.org/10.1016/j.canlet.2023.216424
Br J Pharmacol. 2023 Oct 02.

Transcriptome-level Discovery of Survival-Associated Biomarkers and Therapy Targets in Non-Small-Cell Lung Cancer.

Balázs Győrffy.

   BACKGROUND AND PURPOSE: Survival rate of patients with lung cancer has increased by over 60% in the recent two decades. With longer survival, the identification of genes associated with survival has emerged as an issue of utmost importance to uncover the most promising biomarkers and therapeutic targets.
EXPERIMENTAL APPROACH: An integrated database was set up by combining multiple independent datasets with clinical data and transcriptome-level gene expression measurements. Uni- and multivariate survival analyses were performed to identify genes with higher expression levels linked to shorter survival. The strongest genes were filtered to include only those with known druggability.
KEY RESULTS: The entire database includes 2,852 tumor specimens from 17 independent cohorts. Of these, 2,227 have overall survival data and 1,256 samples have progression-free survival time. The most significant genes associated with survival were MIF, UBC, and B2M in lung adenocarcinoma and ANXA2, CSNK2A2, and KRT18 in squamous cell carcinoma. We also aimed to reveal the best druggable targets in non-smokers lung cancer. The three most promising hits in this cohort were MDK, THY1 and PADI2. The established lung cancer cohort was added to the Kaplan-Meier plotter (https://www.kmplot.com) enabling the validation of future gene expression-based biomarkers in both the present and yet unexamined subgroups of patients.
CONCLUSIONS AND IMPLICATIONS: In this study, we established a comprehensive database of transcriptome-level data for lung cancer. The database can be utilized to identify and rank the most promising biomarkers and therapeutic targets for different subtypes of lung cancer.

Keywords:  cox regression; gene arrays; gene expression; pharmacology; prognosis; survival

DOI:  https://doi.org/10.1111/bph.16257
Int J Radiat Oncol Biol Phys. 2023 Oct 01. pii: S0360-3016(23)05169-6. [Epub ahead of print]117(2S): e40

Prognostic Significance of Serum Lipids in Patients with Non-Small Cell Lung Cancer Treated with Radiotherapy: A Multicenter Prospective Study.

J Lv, T Li, H S Bai, H Kuang, H Jia, C Li, L Liang.

PURPOSE/OBJECTIVE(S): Although lipids have been assessed for their possible roles in cancer survival prediction, studies on the association between serum lipids levels and the prognosis of non-small cell lung cancer (NSCLC) patients are limited. This study aimed to evaluate whether serum lipids are associated with outcomes in patients with NSCLC treated with radiotherapy.
MATERIALS/METHODS: We conducted a multicenter prospective study on patients diagnosed with NSCLC between January 2018 and February 2021. Participants received thoracic radiotherapy of 60ཞ80 Gy to the primary lung tumor and positive lymph node metastases. We measured patients' serum lipids levels (serum triglyceride, TGs; total cholesterol, TC, high density lipoprotein cholesterol, HDL-C; low density lipoprotein cholesterol, LDL-C) before radiotherapy. The association between serum lipids levels and overall survival (OS) was evaluated using hazard ratios. We sought to determine a threshold point using optimal stratification. Survival analysis was performed using Kaplan-Meier curves.
RESULTS: Of the 300 participants diagnosed with NSCLC treated with radiotherapy, 165 (55.0%) were men. Median follow-up time was 24.4 months (range 1.0- 101.9 months). Using univariate and multivariate Cox proportional hazard analysis, among those serum lipids, only serum TG was shown to be independent prognostic factors for OS (hazard ratio: 1.203, 95% confidence interval: 1.038 - 1.393, p = 0.014). The cut-off for TG associated with OS was 2.04 mmol/L. Based on the TG cut-off value, 55 NSCLC patients were categorized into the high TG group (>2.04 mmol/L) and 245 in the low TG group (<2.04 mmol/L). The NSCLC patients in the low TG group exhibited higher OS than the high group (median OS, not reach vs 41.4 months, p = 0.025).
CONCLUSION: TG levels were found to be a significant negative prognostic biomarker for OS in NSCLC patients treated with radiotherapy.

DOI: https://doi.org/10.1016/j.ijrobp.2023.06.735
Int J Radiat Oncol Biol Phys. 2023 Oct 01. pii: S0360-3016(23)05119-2. [Epub ahead of print]117(2S): e17-e18

Prognostic Value of Hypoxia Gene Expression Signature in Early-Stage Non-Small Cell Lung Cancer Patients Treated with Stereotactic Body Radiation Therapy.

N J Eustace, N Sebastian, A Webb, N Denko, K Shilo, R Robb, T M Williams.

PURPOSE/OBJECTIVE(S): Stereotactic body radiation therapy (SBRT) is an effective alternative to surgery in the treatment of early-stage non-small cell lung cancer (NSCLC). However, disease recurrence occurs in approximately 20-30% of patients. Hypoxia is a well-known factor promoting solid tumor progression, radiation resistance, and immune evasion. In this study, we utilize a previously published hypoxia gene expression signature (Buffa) and measure its association with clinical outcomes in patients with NSCLC treated with SBRT.
MATERIALS/METHODS: Our study focused on patients with localized, node-negative NSCLC treated with SBRT, and we identified 92 patients treated at our institution between 2008 and 2018 with available gene expression data. Total RNA from formalin-fixed paraffin-embedded archival biopsy specimens (pre-therapy) was isolated and mRNA expression was analyzed using an assay for human samples. For each gene in the Buffa signature, the top 50% of mRNA abundance values were given a score of +1, and the bottom 50% were given a score of -1 to generate a tumor hypoxia score. High scores suggest a hypoxic tumor and low scores imply normoxia. Kaplan-Meier curves were used to assess overall survival, disease-free survival, local recurrence, regional nodal recurrence, and distant recurrence. Cox proportional hazards were performed to account for confounding due to age, T stage, ECOG performance status, biologically effective dose, patient sex, and histology (squamous vs non-squamous).
RESULTS: The patient's gene expression data were dichotomized based on their median hypoxia score (Table 1). Median follow-up was 23.9 months (95% CI 20.6 - 26.6). A high hypoxia score was associated with squamous histology (p = 0.003). On univariate analysis, a high hypoxia score was significantly associated with local recurrence (hazard ratio [HR] = 5.63; 95% CI 1.03 - 30.78; p = 0.046) and distant recurrence (HR = 1.85; 95% CI 1.14 - 3.02; p = 0.013). There was no significant difference in overall survival (HR = 1.93; 95% CI 0.98 - 3.82; p = 0.058), disease-free survival (HR = 2.13; 95% CI 0.93 - 4.89; p = 0.074), or regional nodal recurrence (HR = 1.45; 95% CI 0.41 - 5.14; p = 0.57). On multivariate analysis, a high hypoxia score was associated with worse overall survival (HR = 2.73; 95% CI 1.23 - 6.07; p = 0.014), local recurrence (HR = 12.85; 95% CI 1.08 - 152.49; p = 0.043) and distant recurrence (HR = 1.91; 95% CI 1.13 - 3.22; p = 0.016).
CONCLUSION: Our findings demonstrate that a hypoxia gene signature is significantly associated with worse survival and increased local and distant recurrence after SBRT for localized NSCLC. This hypoxia signature may be useful in prognostication, selecting patients for surgery versus radiation, or selecting patients for treatment intensification with radiation or other systemic agents, and should be prospectively validated.

DOI: https://doi.org/10.1016/j.ijrobp.2023.06.685
Transl Oncol. 2023 Sep 28. pii: S1936-5233(23)00174-2. [Epub ahead of print]38 101788

Tumor mutation burden in the prognosis and response of lung cancer patients to immune-checkpoint inhibition therapies.

Paraskevi Vryza, Timo Fischer, Elena Mistakidi, Apostolos Zaravinos.

  Immune checkpoint inhibition (ICI) therapies have reshaped the therapeutic landscape in lung cancer management, providing first-time improvements in patient response, prognosis, and overall survival. Despite their clinical effectiveness, variability in treatment responsiveness, as well as drug resistance, have led to a compelling need for predictive biomarkers facilitating the individualized selection of the most efficient therapeutic approach. Significant progress has been made in the identification of such biomarkers, with tumor mutation burden (ΤΜΒ) appearing as the leading and most promising predictive biomarker for the efficacy of ICIs in non-small cell lung cancer (NSCLC) among other tumors. Anti-PD-1/PD-L1 and anti-CTLA-4 antibodies have been extensively studied and clinically utilized. However, the overall efficiency of these drugs remains unsatisfactory, urging for the investigation of novel inhibitors, such as those targeting LAG-3, TIM-3, TIGIT and VISTA, which could be used either as a monotherapy or synergistically with the PD-1/PD-L1 or CTLA-4 blockers. Here, we investigate the role of TMB and cancer neoantigens as predictive biomarkers in the response of lung cancer patients to different ICI therapies, specifically focusing on the most recent immune checkpoint inhibitors, against LAG-3, TIM-3, TIGIT and VISTA. We further discuss the new trends in immunotherapies, including CAR T-cell therapy and personalized tumor vaccines. We also review further potential biomarkers that could be used in lung cancer response to immunotherapy, such as PD-L1+ IHC, MSI/dMMR, tumor infiltrating lymphocytes (TILs), as well as the role of the microbiome and circulating tumor DNA (ctDNA). Finally, we discuss the limitations and challenges of each.

Keywords:  Immune checkpoint inhibition therapy; Patient response; Tumor microenvironment; Tumor mutation burden (TMB); Tumor-infiltrating lymphocytes

DOI:  https://doi.org/10.1016/j.tranon.2023.101788