Differential uptake of antisense oligonucleotides in mouse hepatocytes and macrophages revealed by simultaneous 2P-fluorescence and CARS

Prabuddha Mukherjee; Edita Aksamitiene; Aneesh Alex; Derek Shi; Darold R. Spillman Jr.; Marina Marjanovic; Michael Fazio; Punit Seth; Kendal Frazier; Steve R. Hood; Stephen A. Boppart

doi:10.1117/12.2578037

5 March 2021 Differential uptake of antisense oligonucleotides in mouse hepatocytes and macrophages revealed by simultaneous 2P-fluorescence and CARS

Prabuddha Mukherjee, Edita Aksamitiene, Aneesh Alex, Derek Shi, Darold R. Spillman Jr., Marina Marjanovic, Michael Fazio, Punit Seth, Kendal Frazier, Steve R. Hood, Stephen A. Boppart

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Proceedings Volume 11624, Visualizing and Quantifying Drug Distribution in Tissue V; 116240E (2021) https://doi.org/10.1117/12.2578037
Event: SPIE BiOS, 2021, Online Only

Abstract

Antisense oligonucleotides (ASOs), a novel paradigm in modern therapeutics, modulate cellular gene expression by binding to complementary RNA sequences. Triantennary N-acetyl galactosamine (GN)-conjugated ASOs show greatly improved potency via Asialoglycoprotein receptor (ASGR)-mediated uptake in hepatocytes. Here, we compare the uptake kinetics and subsequent distribution of untargeted ASOs to that of GN-ASOs in mouse macrophages and hepatocytes using simultaneous coherent anti-Stokes Raman scattering (CARS) and two-photon excited fluorescence imaging. While the CARS modality captured the changing lipid distributions and overall morphology of the cell, two-photon fluorescence imaging measured the uptake and the subsequent distribution of the fluorescently labeled (Alexa-488) ASOs inside the cells.

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Prabuddha Mukherjee, Edita Aksamitiene, Aneesh Alex, Derek Shi, Darold R. Spillman Jr., Marina Marjanovic, Michael Fazio, Punit Seth, Kendal Frazier, Steve R. Hood, and Stephen A. Boppart "Differential uptake of antisense oligonucleotides in mouse hepatocytes and macrophages revealed by simultaneous 2P-fluorescence and CARS", Proc. SPIE 11624, Visualizing and Quantifying Drug Distribution in Tissue V, 116240E (5 March 2021); https://doi.org/10.1117/12.2578037

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