Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

Keynote webinar | Spotlight on medication adherence

LIVE: Thursday 27th June 2024, 18:00-19:30 (CEST)
Join our expert panel to discover why you need to understand the drivers of non-adherence in your patients, and how you can optimize medication adherence in your clinics to drastically improve patient outcomes.
ADVANCED SEARCH
Log in
Top
Cancer Cell International
Published in: Cancer Cell International 1/2020

01-12-2020 | Addiction | Primary research

Palbociclib treatment alters nucleotide biosynthesis and glutamine dependency in A549 cells

Authors: Lindsey R. Conroy, Pawel Lorkiewicz, Liqing He, Xinmin Yin, Xiang Zhang, Shesh N. Rai, Brian F. Clem

Published in: Cancer Cell International | Issue 1/2020

Login to get access

Abstract

Background

Aberrant activity of cell cycle proteins is one of the key somatic events in non-small cell lung cancer (NSCLC) pathogenesis. In most NSCLC cases, the retinoblastoma protein tumor suppressor (RB) becomes inactivated via constitutive phosphorylation by cyclin dependent kinase (CDK) 4/6, leading to uncontrolled cell proliferation. Palbociclib, a small molecule inhibitor of CDK4/6, has shown anti-tumor activity in vitro and in vivo, with recent studies demonstrating a functional role for palbociclib in reprogramming cellular metabolism. While palbociclib has shown efficacy in preclinical models of NSCLC, the metabolic consequences of CDK4/6 inhibition in this context are largely unknown.

Methods

In our study, we used a combination of stable isotope resolved metabolomics using [U-13C]-glucose and multiple in vitro metabolic assays, to interrogate the metabolic perturbations induced by palbociclib in A549 lung adenocarcinoma cells. Specifically, we assessed changes in glycolytic activity, the pentose phosphate pathway (PPP), and glutamine utilization. We performed these studies following palbociclib treatment with simultaneous silencing of RB1 to define the pRB-dependent changes in metabolism.

Results

Our studies revealed palbociclib does not affect glycolytic activity in A549 cells but decreases glucose metabolism through the PPP. This is in part via reducing activity of glucose 6-phosphate dehydrogenase, the rate limiting enzyme in the PPP. Additionally, palbociclib enhances glutaminolysis to maintain mitochondrial respiration and sensitizes A549 cells to the glutaminase inhibitor, CB-839. Notably, the effects of palbociclib on both the PPP and glutamine utilization occur in an RB-dependent manner.

Conclusions

Together, our data define the metabolic impact of palbociclib treatment in A549 cells and may support the targeting CDK4/6 inhibition in combination with glutaminase inhibitors in NSCLC patients with RB-proficient tumors.
Appendix
Available only for authorised users
Literature
1.
2.
Gridelli C, Rossi A, Carbone DP, Guarize J, Karachaliou N, Mok T, et al. Non-small-cell lung cancer. Nat Rev Dis Primers. 2015;1:15009.PubMedCrossRef
3.
Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J, Jemal A. Global cancer statistics, 2012. CA Cancer J Clin. 2015;65(2):87–108.CrossRef
4.
Esposito V, Baldi A, Tonini G, Vincenzi B, Santini M, Ambrogi V, Mineo TC, Persichetti P, Liuzzi G, Montesarchio V, Wolner E, Baldi F, Groeger AM. Analysis of cell cycle regulator proteins in non-small cell lung cancer. J Clin Pathol. 2004;57:58–63.PubMedPubMedCentralCrossRef
5.
Eymin B, Gazzeri S. Role of cell cycle regulators in lung carcinogenesis. Cell Adhes Migrat. 2009;4:114–23.CrossRef
6.
7.
Klein ME, Kovatcheva M, Davis LE, Tap WD, Koff A. CDK4/6 inhibitors: the mechanism of action may not be as simple as once thought. Cancer Cell. 2018;34(1):9–20.PubMedPubMedCentralCrossRef
8.
Beaver JA, Amiri-Kordestani L, Charlab R, Chen W, Palmby T, Tilley A, et al. FDA approval: palbociclib for the treatment of postmenopausal patients with estrogen receptor-positive, HER2-negative metastatic breast cancer. Clin Cancer Res. 2015;21(21):4760–6.PubMedCrossRef
9.
Gopalan PK, Villegas AG, Cao C, Pinder-Schenck M, Chiappori A, Hou W, et al. CDK4/6 inhibition stabilizes disease in patients with p16-null non-small cell lung cancer and is synergistic with mTOR inhibition. Oncotarget. 2018;9(100):37352–66.PubMedPubMedCentralCrossRef
10.
Liu M, Xu S, Wang Y, Li Y, Li Y, Zhang H, et al. PD 0332991, a selective cyclin D kinase 4/6 inhibitor, sensitizes lung cancer cells to treatment with epidermal growth factor receptor tyrosine kinase inhibitors. Oncotarget. 2016;7(51):84951–64.PubMedPubMedCentralCrossRef
11.
Nie H, Zhou X, Shuzhang D, Nie C, Zhang X, Huang J. Palbociclib overcomes afatinib resistance in non-small cell lung cancer. Biomed Pharmacother. 2019;109:1750–7.PubMedCrossRef
12.
13.
Zhou J, Zhang S, Chen X, Zheng X, Yao Y, Lu G, et al. Palbociclib, a selective CDK4/6 inhibitor, enhances the effect of selumetinib in RAS-driven non-small cell lung cancer. Cancer Lett. 2017;408:130–7.PubMedCrossRef
14.
Franco J, Balaji U, Freinkman E, Witkiewicz AK, Knudsen ES. Metabolic reprogramming of pancreatic cancer mediated by CDK4/6 inhibition elicits unique vulnerabilities. Cell Rep. 2016;14(5):979–90.PubMedPubMedCentralCrossRef
15.
Tarrado-Castellarnau M, de Atauri P, Tarrago-Celada J, Perarnau J, Yuneva M, Thomson TM, et al. De novo MYC addiction as an adaptive response of cancer cells to CDK4/6 inhibition. Mol Syst Biol. 2017;13(10):940.PubMedPubMedCentralCrossRef
16.
Wang H, Nicolay BN, Chick JM, Gao X, Geng Y, Ren H, et al. The metabolic function of cyclin D3-CDK6 kinase in cancer cell survival. Nature. 2017;546(7658):426–30.PubMedPubMedCentralCrossRef
17.
Klavins K, Drexler H, Hann S, Koellensperger G. Quantitative metabolite profiling utilizing parallel column analysis for simultaneous reversed-phase and hydrophilic interaction liquid chromatography separations combined with tandem mass spectrometry. Anal Chem. 2014;86(9):4145–50.PubMedCrossRef
18.
Adusumilli R, Mallick P. Data conversion with ProteoWizard msConvert. Methods Mol Biol. 2017;1550:339–68.PubMedCrossRef
19.
Agrawal S, Kumar S, Sehgal R, George S, Gupta R, Poddar S, et al. El-MAVEN: a fast, robust, and user-friendly mass spectrometry data processing engine for metabolomics. In: D’Alessandro A, editor. High-throughput metabolomics: methods and protocols. 1978th ed. New York: Springer; 2019.
20.
Salabei JK, Lorkiewicz PK, Mehra P, Gibb AA, Haberzettl P, Hong KU, et al. Type 2 diabetes dysregulates glucose metabolism in cardiac progenitor Cells. J Biol Chem. 2016;291(26):13634–48.PubMedPubMedCentralCrossRef
21.
Wei X, Lorkiewicz PK, Shi B, Salabei JK, Hill BG, Kim S, et al. Analysis of stable isotope assisted metabolomics data acquired by high resolution mass spectrometry. Anal Methods. 2017;9:2275–83.PubMedPubMedCentralCrossRef
22.
Lorkiewicz PK, Gibb AA, Rood BR, He L, Zheng Y, Clem BF, et al. Integration of flux measurements and pharmacological controls to optimize stable isotope-resolved metabolomics workflows and interpretation. Scientific Reports. 2019;9(1):1–7.CrossRef
23.
Cretella D, Fumarola C, Bonelli M, Alfieri R, La Monica S, Digiacomo G, et al. Pre-treatment with the CDK4/6 inhibitor palbociclib improves the efficacy of paclitaxel in TNBC cells. Sci Rep. 2019;9(1):13014.PubMedPubMedCentralCrossRef
24.
Cretella D, Ravelli A, Fumarola C, La Monica S, Digiacomo G, Cavazzoni A, et al. The anti-tumor efficacy of CDK4/6 inhibition is enhanced by the combination with PI3K/AKT/mTOR inhibitors through impairment of glucose metabolism in TNBC cells. J Exp Clin Cancer Res. 2018;37(1):72.PubMedPubMedCentralCrossRef
25.
Knudsen ES, Witkiewicz AK. Defining the transcriptional and biological response to CDK4/6 inhibition in relation to ER+/HER2− breast cancer. Oncotarget. 2016;7(43):69111–23.PubMedPubMedCentralCrossRef
26.
Daye D, Wellen KE. Metabolic reprogramming in cancer: unraveling the role of glutamine in tumorigenesis. Semin Cell Dev Biol. 2012;23(4):362–9.PubMedCrossRef
27.
DeBerardinis RJ, Cheng T. Q’s next: the diverse functions of glutamine in metabolism, cell biology and cancer. Oncogene. 2010;29(3):313–24.PubMedCrossRef
28.
DeBerardinis RJ, Mancuso A, Daikhin E, Nissim I, Yudkoff M, Wehrli S, et al. Beyond aerobic glycolysis: transformed cells can engage in glutamine metabolism that exceeds the requirement for protein and nucleotide synthesis. Proc Natl Acad Sci USA. 2007;104(49):19345–50.PubMedCrossRef
29.
Boysen G, Jamshidi-Parsian A, Davis MA, Siegel ER, Simecka CM, Kore RA, et al. Glutaminase inhibitor CB-839 increases radiation sensitivity of lung tumor cells and human lung tumor xenografts in mice. Int J Radiat Biol. 2019;95(4):436–42.PubMedPubMedCentralCrossRef
30.
Cairns RA, Harris IS, Mak TW. Regulation of cancer cell metabolism. Nat Rev Cancer. 2011;11(2):85–95.PubMedCrossRef
31.
32.
DeBerardinis RJ, Lum JJ, Hatzivassiliou G, Thompson CB. The biology of cancer: metabolic reprogramming fuels cell growth and proliferation. Cell Metab. 2008;7(1):11–20.PubMedCrossRef
33.
Martinez-Outschoorn UE, Peiris-Pages M, Pestell RG, Sotgia F, Lisanti MP. Cancer metabolism: a therapeutic perspective. Nat Rev Clin Oncol. 2017;14(1):11–31.PubMedCrossRef
34.
35.
Blanchet E, Annicotte JS, Lagarrigue S, Aguilar V, Clape C, Chavey C, et al. E2F transcription factor-1 regulates oxidative metabolism. Nat Cell Biol. 2011;13(9):1146–52.PubMedCrossRef
36.
Bonelli M, La Monica S, Fumarola C, Alfieri R. Multiple effects of CDK4/6 inhibition in cancer: from cell cycle arrest to immunomodulation. Biochem Pharmacol. 2019;170:113676.PubMedCrossRef
37.
38.
Clem BF, Chesney J. Molecular pathways: regulation of metabolism by RB. Clin Cancer Res. 2012;18(22):6096–100.PubMedCrossRef
39.
Gao P, Tchernyshyov I, Chang TC, Lee YS, Kita K, Ochi T, et al. c-Myc suppression of miR-23a/b enhances mitochondrial glutaminase expression and glutamine metabolism. Nature. 2009;458(7239):762–5.PubMedPubMedCentralCrossRef
40.
Goetzman ES, Prochownik EV. The role for Myc in coordinating glycolysis, oxidative phosphorylation, glutaminolysis, and fatty acid metabolism in normal and neoplastic tissues. Front Endocrinol. 2018;9:129.CrossRef
41.
Takebayashi S, Tanaka H, Hino S, Nakatsu Y, Igata T, Sakamoto A, et al. Retinoblastoma protein promotes oxidative phosphorylation through upregulation of glycolytic genes in oncogene-induced senescent cells. Aging Cell. 2015;14(4):689–97.PubMedPubMedCentralCrossRef
42.
Thangavel C, Boopathi E, Liu Y, McNair C, Haber A, Perepelyuk M, et al. Therapeutic challenge with a CDK 4/6 inhibitor induces an RB-dependent SMAC-mediated apoptotic response in non-small cell lung cancer. Clin Cancer Res. 2018;24:1402–14.PubMedPubMedCentralCrossRef
43.
Vijayaraghavan S, Karakas C, Doostan I, Chen X, Bui T, Yi M, et al. CDK4/6 and autophagy inhibitors synergistically induce senescence in Rb positive cytoplasmic cyclin E negative cancers. Nat Commun. 2017;8:15916.PubMedPubMedCentralCrossRef
44.
45.
46.
Metadata
Title
Palbociclib treatment alters nucleotide biosynthesis and glutamine dependency in A549 cells
Authors
Lindsey R. Conroy
Pawel Lorkiewicz
Liqing He
Xinmin Yin
Xiang Zhang
Shesh N. Rai
Brian F. Clem
Publication date
01-12-2020
Publisher
BioMed Central
Published in
Cancer Cell International / Issue 1/2020
Electronic ISSN: 1475-2867
DOI
https://doi.org/10.1186/s12935-020-01357-x

Other articles of this Issue 1/2020

Cancer Cell International 1/2020 Go to the issue

Primary research

LncRNA ST7-AS1, by regulating miR-181b-5p/KPNA4 axis, promotes the malignancy of lung adenocarcinoma

Primary research

Circular RNA ATXN7 promotes the development of gastric cancer through sponging miR-4319 and regulating ENTPD4

Primary research

Down-regulation of TRIB3 inhibits the progression of ovarian cancer via MEK/ERK signaling pathway

Primary research

Comprehensive analysis of LDHAP5 pseudogene expression and potential pathogenesis in ovarian serous cystadenocarcinoma

Primary research

Kinesin superfamily protein 21B acts as an oncogene in non-small cell lung cancer

Primary research

A robust twelve-gene signature for prognosis prediction of hepatocellular carcinoma
Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras
Prof. Fabrice Barlesi
Developed by: Springer Medicine
Image Credits