June 2022
Volume 63, Issue 7
Open Access
ARVO Annual Meeting Abstract  |   June 2022
PLL-g-PEG inhibits antibody-drug conjugate uptake into human corneal epithelial cells in vitro.
Author Affiliations & Notes
  • David Kleinman
    Calm Water Therapeutics LLC, Rochester, New York, United States
  • Jenni J. Hakkarainen
    R&D Division, Experimentica Ltd., Kuopio, Finland
  • Anita Kirti Ghosh
    R&D Division, Experimentica Ltd., Forest Park, Illinois, United States
  • Sean David Ogle
    R&D Division, Experimentica Ltd., Forest Park, Illinois, United States
  • Kevin Sill
    Calm Water Therapeutics LLC, Rochester, New York, United States
  • Brian A Mendelsohn
    Exelixis, Alameda, California, United States
  • Mark Mitchnick
    Calm Water Therapeutics LLC, Rochester, New York, United States
  • Simon Kaja
    R&D Division, Experimentica Ltd., Forest Park, Illinois, United States
    Departments of Ophthalmology and Molecular Pharmacology & Neuroscience, Loyola University Chicago, Chicago, Illinois, United States
  • Footnotes
    Commercial Relationships   David Kleinman AGTC, Aprea Therapeutics, Cleave Therapeutics, Coherus BioSciences, Design Therapeutics, Editas Medicine, GSK, Helixmith USA, Kala Pharmaceuticals, Olema Pharmaceuticals, ONL Therapeutics, Revolution Medicines, Triphase Accelerator, Code C (Consultant/Contractor), Calm Water Therapeutics LLC, ONL Therapeutics, Inc., Code I (Personal Financial Interest), Calm Water Therapeutics, Code O (Owner), Calm Water Therapeutics, LLC, Code S (non-remunerative); Jenni Hakkarainen Experimentica Ltd., Code E (Employment), Experimentica Ltd., Code I (Personal Financial Interest), Experimentica Ltd., Code S (non-remunerative); Anita Ghosh Experimentica Ltd., K&P Scientific LLC, Code C (Consultant/Contractor), Experimentica Ltd., Code E (Employment), eyeNOS Inc., Code I (Personal Financial Interest), eyeNOS Inc., Code P (Patent), Experimentica Ltd., K&P Scientific LLC, Code R (Recipient), Experimentica Ltd., eyeNOS Inc., Code S (non-remunerative); Sean Ogle eyeNOS Inc., Code C (Consultant/Contractor), Experimentica Ltd., Code E (Employment); Kevin Sill Calm Water Therapeutics LLC, Code C (Consultant/Contractor), Calm Water Therapeutics LLC, Code I (Personal Financial Interest); Brian Mendelsohn Exelixis, Code E (Employment), Exelixis, Code I (Personal Financial Interest); Mark Mitchnick Calm Water Therapeutics LLC, Code I (Personal Financial Interest), Calm Water Therapeutics LLC, Code S (non-remunerative); Simon Kaja Experimentica Ltd., Code C (Consultant/Contractor), Experimentica Ltd., K&P Scientific LLC , Code F (Financial Support), Experimentica Ltd., K&P Scientific LLC , Code I (Personal Financial Interest), eyeNOS Inc., Code P (Patent), Experimentica Ltd., K&P Scientific LLC, Code R (Recipient), Experimentica Ltd., K&P Scientific LLC , Code S (non-remunerative)
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science June 2022, Vol.63, 3239 – A0274. doi:
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    • Get Citation

      David Kleinman, Jenni J. Hakkarainen, Anita Kirti Ghosh, Sean David Ogle, Kevin Sill, Brian A Mendelsohn, Mark Mitchnick, Simon Kaja; PLL-g-PEG inhibits antibody-drug conjugate uptake into human corneal epithelial cells in vitro.. Invest. Ophthalmol. Vis. Sci. 2022;63(7):3239 – A0274.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Corneal toxicity secondary to antibody-drug conjugates (ADCs) is a common and clinically significant adverse event that interferes with ADC dosing and could ultimately affect a patient's response to ADC therapy. Effective interventions that either prevent or mitigate this drug-induced keratopathy are lacking. Evidence suggests that ADCs can enter corneal epithelial cells or their precursors indirectly through macropinocytosis, often with no involvement of the targeted receptor. Once internalized, the drug is cleaved from the ADC releasing a cytotoxic small molecule. This present study was based on the rationale that preventing the uptake of an ADC into off-target tissue should mitigate toxicity and the knowledge that charge plays an important role in the interaction of immunoconjugates with cell membranes. The hypothesis that locally delivered poly(L-lysine)-g-poly(ethylene glycol) (PLL-g-PEG) can inhibit ADC uptake into human corneal epithelial cells was tested.

Methods : Rituximab-monomethyl auristatin F (MMAF) was selected as an exemplary ADC that will demonstrate off target corneal epithelial cell toxicity. PLL (not PLL-g-PEG) - coated chamber slides were seeded with human corneal epithelial - transformed (HCE-T) cells (50,000 cells/cm2). After 48 hours, cells were treated with test article (PLL-g-PEG, or positive control, 5-(N-ethyl-Nisopropyl) amiloride (EIPA)) or vehicle. After 30 minutes, Rituximab-MMAF (75 nM) was added to the wells. The wells were maintained for an additional 3.5 hours before HCE-T cells were fixed and ADC was detected by fluorescent anti-human IgG as viewed under microscopy. Fluorescence intensity was reported utilizing multiple images obtained from multiple replicates. A pilot evaluation was carried out, followed by two validation studies.

Results : The pilot evaluation demonstrated a statistically significant reduction in fluorescence intensity when vehicle was compared to EIPA (150 µmol/L) and PLL-g-PEG 1% solution. Two validation studies demonstrated a dose dependent decrease in fluorescence compared to vehicle for PLL-g-PEG concentrations of 0.05%, 0.1%, 0.5%, and 1% (see images).

Conclusions : PLL-g-PEG exhibited a dose-dependent inhibition of ADC uptake into HCE-T cells. These results suggest PLL-g-PEG may be an effective topical therapy to prevent, mitigate, or otherwise reduce ADC-induced corneal epithelial cell toxicity.

This abstract was presented at the 2022 ARVO Annual Meeting, held in Denver, CO, May 1-4, 2022, and virtually.

 

Anti-IgG Fluoresence Intensity.

Anti-IgG Fluoresence Intensity.

 

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