Inhibition of EGF Uptake by Nephrotoxic Antisense Drugs In Vitro and Implications for Preclinical Safety Profiling.
Antisense oligonucleotide (AON) therapeutics offer new avenues to pursue clinically relevant targets inaccessible with other technologies. Advances in improving AON affinity and stability by incorporation of high affinity nucleotides, such as locked nucleic acids (LNA), have sometimes been stifled by safety liabilities related to their accumulation in the kidney tubule. In an attempt to predict and understand the mechanisms of LNA-AON-induced renal tubular toxicity, we established human cell models that recapitulate in vivo behavior of pre-clinically and clinically unfavorable LNA-AON drug candidates. We identified elevation of extracellular epidermal growth factor (EGF) as a robust and sensitive in vitro biomarker of LNA-AON-induced cytotoxicity in human kidney tubule epithelial cells. We report the time-dependent negative regulation of EGF uptake and EGF receptor (EGFR) signaling by toxic but not innocuous LNA-AONs and revealed the importance of EGFR signaling in LNA-AON-mediated decrease in cellular activity. The robust EGF-based in vitro safety profiling of LNA-AON drug candidates presented here, together with a better understanding of the underlying molecular mechanisms, constitutes a significant step toward developing safer antisense therapeutics.
Roche Pharma Research and Early Development, Roche Innovation Center Basel, Basel 4070, Switzerland. Electronic address: annie.moisan@roche.com.
Annie
A
Moisan
Roche Pharma Research and Early Development, Roche Innovation Center Basel, Basel 4070, Switzerland.
Marcel
M
Gubler
Roche Pharma Research and Early Development, Roche Innovation Center Basel, Basel 4070, Switzerland.
Jitao David
JD
Zhang
Roche Pharma Research and Early Development, Roche Innovation Center Copenhagen, Hørsholm 2970, Denmark.
Yann
Y
Tessier
Roche Pharma Research and Early Development, Roche Innovation Center Copenhagen, Hørsholm 2970, Denmark.
Kamille
K
Dumong Erichsen
Roche Pharma Research and Early Development, Roche Innovation Center Basel, Basel 4070, Switzerland.
Sabine
S
Sewing
Roche Pharma Research and Early Development, Roche Innovation Center Basel, Basel 4070, Switzerland.
Régine
R
Gérard
Roche Pharma Research and Early Development, Roche Innovation Center Basel, Basel 4070, Switzerland.
Blandine
B
Avignon
Roche Pharma Research and Early Development, Roche Innovation Center Basel, Basel 4070, Switzerland.
Sylwia
S
Huber
Roche Pharma Research and Early Development, Roche Innovation Center Basel, Basel 4070, Switzerland.
Fethallah
F
Benmansour
Roche Pharma Research and Early Development, Roche Innovation Center Basel, Basel 4070, Switzerland.
Xing
X
Chen
Roche Pharma Research and Early Development, Roche Innovation Center Basel, Basel 4070, Switzerland.
Roberto
R
Villaseñor
Roche Pharma Research and Early Development, Roche Innovation Center Basel, Basel 4070, Switzerland.
Annamaria
A
Braendli-Baiocco
Roche Pharma Research and Early Development, Roche Innovation Center Basel, Basel 4070, Switzerland.
Matthias
M
Festag
Roche Pharma Research and Early Development, Roche Innovation Center Basel, Basel 4070, Switzerland.
Andreas
A
Maunz
Roche Pharma Research and Early Development, Roche Innovation Center Basel, Basel 4070, Switzerland.
Thomas
T
Singer
Roche Pharma Research and Early Development, Roche Innovation Center Basel, Basel 4070, Switzerland.
Franz
F
Schuler
Roche Pharma Research and Early Development, Roche Innovation Center Basel, Basel 4070, Switzerland.
Adrian B
AB
Roth
EGF; EGFR; PTEC; antisense; kidney; nephrotoxicity; oligonucleotide; preclinical; safety