TY - JOUR
T1 - Anti-glioblastoma activity of monensin and its analogs in an organoid model of cancer
AU - Urbaniak, Alicja
AU - Reed, Megan R.
AU - Heflin, Billie
AU - Gaydos, John
AU - Piña-Oviedo, Sergio
AU - Jędrzejczyk, Marta
AU - Klejborowska, Greta
AU - Stępczyńska, Natalia
AU - Chambers, Timothy C.
AU - Tackett, Alan J.
AU - Rodriguez, Analiz
AU - Huczyński, Adam
AU - Eoff, Robert L.
AU - MacNicol, Angus M.
N1 - Funding Information:
This work was supported by Winthrop P. Rockefeller Cancer Institute Team Science Award (to AMM, RLE and AR), a Barton Pilot Grant from UAMS College of Medicine (to AU), Equipment Award Program from UAMS College of Medicine (to AU), and grants from the National Institutes of Health: R01HD093461 (to AMM), P20GM121293 (to AJT), R24GM137786 (to AJT), and R01CA236209 (to AJT). This publication was also supported by Grant Number P30 GM103450 from the National Institute of General Medical Sciences of the National Institutes of Health (NIH). We thank Ms. Andrea Harris from the UAMS Flow Cytometry Core, Ms. Jennifer James from the UAMS Experimental Pathology Core and Mr. Jeff Kamykowski from the UAMS Digital Microscopy Core Laboratory. We would like to thank Drs. Eric Norris and HoWon Kim from StemCell Technologies for valuable discussions concerning iPSC culture and COs generation. We also thank Dr. Amanda Linkous for consultations and helpful suggestions for the generation of the GLICO model. Some elements in the figures were created using BioRender (BioRender.com).
Funding Information:
This work was supported by Winthrop P. Rockefeller Cancer Institute Team Science Award (to AMM, RLE and AR), a Barton Pilot Grant from UAMS College of Medicine (to AU), Equipment Award Program from UAMS College of Medicine (to AU), and grants from the National Institutes of Health : R01HD093461 (to AMM), P20GM121293 (to AJT), R24GM137786 (to AJT), and R01CA236209 (to AJT). This publication was also supported by Grant Number P30 GM103450 from the National Institute of General Medical Sciences of the National Institutes of Health (NIH). We thank Ms. Andrea Harris from the UAMS Flow Cytometry Core, Ms. Jennifer James from the UAMS Experimental Pathology Core and Mr. Jeff Kamykowski from the UAMS Digital Microscopy Core Laboratory. We would like to thank Drs. Eric Norris and HoWon Kim from StemCell Technologies for valuable discussions concerning iPSC culture and COs generation. We also thank Dr. Amanda Linkous for consultations and helpful suggestions for the generation of the GLICO model. Some elements in the figures were created using BioRender ( BioRender.com ).
Publisher Copyright:
© 2022 The Authors
PY - 2022/9
Y1 - 2022/9
N2 - Glioblastoma (GBM) remains the most frequently diagnosed primary malignant brain cancer in adults. Despite recent progress in understanding the biology of GBM, the clinical outcome for patients remains poor, with a median survival of approximately one year after diagnosis. One factor contributing to failure in clinical trials is the fact that traditional models used in GBM drug discovery poorly recapitulate patient tumors. Previous studies have shown that monensin (MON) analogs, namely esters and amides on C-26 were potent towards various types of cancer cell lines. In the present study we have investigated the activity of these molecules in GBM organoids, as well as in a host:tumor organoid model. Using a mini-ring cell viability assay we have identified seven analogs (IC50 = 91.5 ± 54.4–291.7 ± 68.8 nM) more potent than parent MON (IC50 = 612.6 ± 184.4 nM). Five of these compounds induced substantial DNA fragmentation in GBM organoids, suggestive of apoptotic cell death. The most active analog, compound 1, significantly reduced GBM cell migration, induced PARP degradation, diminished phosphorylation of STAT3, Akt and GSK3β, increased ɣH2AX signaling and upregulated expression of the autophagy associated marker LC3-II. To investigate the activity of MON and compound 1 in a tumor microenvironment, we developed human cerebral organoids (COs) from human induced pluripotent stem cells (iPSCs). The COs showed features of early developing brain such as multiple neural rosettes with a proliferative zone of neural stem cells (Nestin+), neurons (TUJ1 +), primitive ventricular system (SOX2 +/Ki67 +), intermediate zone (TBR2 +) and cortical plate (MAP2 +). In order to generate host:tumor organoids, we co-cultured RFP-labeled U87MG cells with fully formed COs. Compound 1 and MON reduced U87MG tumor size in the COs after four days of treatment and induced a significant reduction of PARP expression. These findings highlight the therapeutic potential of MON analogs towards GBM and support the application of organoid models in anti-cancer drug discovery.
AB - Glioblastoma (GBM) remains the most frequently diagnosed primary malignant brain cancer in adults. Despite recent progress in understanding the biology of GBM, the clinical outcome for patients remains poor, with a median survival of approximately one year after diagnosis. One factor contributing to failure in clinical trials is the fact that traditional models used in GBM drug discovery poorly recapitulate patient tumors. Previous studies have shown that monensin (MON) analogs, namely esters and amides on C-26 were potent towards various types of cancer cell lines. In the present study we have investigated the activity of these molecules in GBM organoids, as well as in a host:tumor organoid model. Using a mini-ring cell viability assay we have identified seven analogs (IC50 = 91.5 ± 54.4–291.7 ± 68.8 nM) more potent than parent MON (IC50 = 612.6 ± 184.4 nM). Five of these compounds induced substantial DNA fragmentation in GBM organoids, suggestive of apoptotic cell death. The most active analog, compound 1, significantly reduced GBM cell migration, induced PARP degradation, diminished phosphorylation of STAT3, Akt and GSK3β, increased ɣH2AX signaling and upregulated expression of the autophagy associated marker LC3-II. To investigate the activity of MON and compound 1 in a tumor microenvironment, we developed human cerebral organoids (COs) from human induced pluripotent stem cells (iPSCs). The COs showed features of early developing brain such as multiple neural rosettes with a proliferative zone of neural stem cells (Nestin+), neurons (TUJ1 +), primitive ventricular system (SOX2 +/Ki67 +), intermediate zone (TBR2 +) and cortical plate (MAP2 +). In order to generate host:tumor organoids, we co-cultured RFP-labeled U87MG cells with fully formed COs. Compound 1 and MON reduced U87MG tumor size in the COs after four days of treatment and induced a significant reduction of PARP expression. These findings highlight the therapeutic potential of MON analogs towards GBM and support the application of organoid models in anti-cancer drug discovery.
KW - Analogs
KW - Glioblastoma
KW - Monensin
KW - Organoid
UR - http://www.scopus.com/inward/record.url?scp=85134794448&partnerID=8YFLogxK
U2 - 10.1016/j.biopha.2022.113440
DO - 10.1016/j.biopha.2022.113440
M3 - Article
C2 - 36076555
AN - SCOPUS:85134794448
SN - 0753-3322
VL - 153
JO - Biomedicine and Pharmacotherapy
JF - Biomedicine and Pharmacotherapy
M1 - 113440
ER -