TY - JOUR
T1 - A ferrocene-containing nucleoside analogue targets DNA replication in pancreatic cancer cells
AU - Rana, Marium
AU - Perotti, Alessio
AU - Bisset, Lucy M
AU - Smith, James D
AU - Lamden, Emma
AU - Khan, Zahra
AU - Ismail, Media K
AU - Ellis, Katherine
AU - Armstrong, Katie A
AU - Hodder, Samantha L
AU - Bertoli, Cosetta
AU - Meneguello, Leticia
AU - de Bruin, Robertus A M
AU - Morris, Joanna R
AU - Romero-Canelon, Isolda
AU - Tucker, James H R
AU - Hodges, Nikolas J
N1 - © The Author(s) 2022. Published by Oxford University Press.
PY - 2022/6/11
Y1 - 2022/6/11
N2 - Pancreatic ductal adenocarcinoma (PDAC) is a disease that remains refractory to existing treatments including the nucleoside analogue gemcitabine. In the current study we demonstrate that an organometallic nucleoside analogue, the ferronucleoside 1-(S, Rp), is cytotoxic in a panel of PDAC cell lines including gemcitabine resistant MIAPaCa2, with IC50 values comparable to cisplatin. Biochemical studies show that the mechanism of action is inhibition of DNA-replication, S-phase cell cycle arrest and stalling of DNA-replication forks which were directly observed at single molecule resolution by DNA-fibre fluorography. In agreement with this, transcriptional changes following treatment with 1-(S, Rp) include activation of three of the four genes (HUS1, RAD1, RAD17) of the 9-1-1 check point complex clamp and two of the three genes (MRE11, NBN) that form the MRN complex as well as activation of multiple downstream targets. Furthermore, there was evidence of phosphorylation of checkpoint kinases 1 and 2 as well as RPA1 and gamma H2AX, all of which are considered biochemical markers of replication stress. Studies in p53 deficient cell lines showed activation of CDKN1A (p21) and GADD45A by 1-(S, Rp) was at least partially independent of p53. In conclusion, because of its potency and activity in gemcitabine resistant cells, 1-(S, Rp) is a promising candidate molecule for development of new treatments for PDAC.
AB - Pancreatic ductal adenocarcinoma (PDAC) is a disease that remains refractory to existing treatments including the nucleoside analogue gemcitabine. In the current study we demonstrate that an organometallic nucleoside analogue, the ferronucleoside 1-(S, Rp), is cytotoxic in a panel of PDAC cell lines including gemcitabine resistant MIAPaCa2, with IC50 values comparable to cisplatin. Biochemical studies show that the mechanism of action is inhibition of DNA-replication, S-phase cell cycle arrest and stalling of DNA-replication forks which were directly observed at single molecule resolution by DNA-fibre fluorography. In agreement with this, transcriptional changes following treatment with 1-(S, Rp) include activation of three of the four genes (HUS1, RAD1, RAD17) of the 9-1-1 check point complex clamp and two of the three genes (MRE11, NBN) that form the MRN complex as well as activation of multiple downstream targets. Furthermore, there was evidence of phosphorylation of checkpoint kinases 1 and 2 as well as RPA1 and gamma H2AX, all of which are considered biochemical markers of replication stress. Studies in p53 deficient cell lines showed activation of CDKN1A (p21) and GADD45A by 1-(S, Rp) was at least partially independent of p53. In conclusion, because of its potency and activity in gemcitabine resistant cells, 1-(S, Rp) is a promising candidate molecule for development of new treatments for PDAC.
KW - Ferrocene, DNA replication, nucleoside analogue, replication fork arrest, pancreatic cancer
U2 - 10.1093/mtomcs/mfac041
DO - 10.1093/mtomcs/mfac041
M3 - Article
C2 - 35689667
JO - Metallomics
JF - Metallomics
SN - 1756-5901
ER -
