Dual bioluminescence and near-infrared fluorescence monitoring to evaluate spherical nucleic acid nanoconjugate activity in vivo

Timothy L. Sita, Foteini Kouri, Lisa A. Hurley, Timothy J. Merkel, Alexandra Chalastanis, Jasmine L. May, Serena T. Ghelfi, Lisa E. Cole, Thomas C. Cayton, Stacey N. Barnaby, Anthony J. Sprangers, Nikunjkumar Savalia, Charles David James, Andrew Lee, Chad A Mirkin*, Alexander H Stegh

*Corresponding author for this work

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

RNA interference (RNAi)-based gene regulation platforms have shown promise as a novel class of therapeutics for the precision treatment of cancer. Techniques in preclinical evaluation of RNAibased nanoconjugates have yet to allow for optimization of their gene regulatory activity.We have developed spherical nucleic acids (SNAs) as a blood-brain barrier-/blood-tumor barrier-penetrating nanoconjugate to deliver small interfering (si) and micro (mi)RNAs to intracranial glioblastoma (GBM) tumor sites. To identify high-activity SNA conjugates and to determine optimal SNA treatment regimens, we developed a reporter xenograft model to evaluate SNA efficacy in vivo. Engrafted tumors stably coexpress optical reporters for luciferase and a nearinfrared (NIR) fluorescent protein (iRFP670), with the latter fused to the DNA repair protein O6-methylguanine-DNA-methyltransferase (MGMT). Using noninvasive imaging of animal subjects bearing reporter-modified intracranial xenografts, we quantitatively assessed MGMT knockdown by SNAs composed of MGMT-targeting siRNA duplexes (siMGMT-SNAs). We show that systemic administration of siMGMT-SNAs via single tail vein injection is capable of robust intratumoral MGMT protein knockdown in vivo, with persistent and SNA dose-dependent MGMT silencing confirmed by Western blotting of tumor tissue ex vivo. Analyses of SNA biodistribution and pharmacokinetics revealed rapid intratumoral uptake and significant intratumoral retention that increased the antitumor activity of coadministered temozolomide (TMZ). Our study demonstrates that dual noninvasive bioluminescence and NIR fluorescence imaging of cancer xenograft models represents a powerful in vivo strategy to identify RNAi-based nanotherapeutics with potent gene silencing activity and will inform additional preclinical and clinical investigations of these constructs.

Original languageEnglish (US)
Pages (from-to)4129-4134
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume114
Issue number16
DOIs
StatePublished - Apr 18 2017

Fingerprint

Nanoconjugates
Nucleic Acids
Fluorescence
Methyltransferases
Heterografts
DNA
Neoplasms
temozolomide
RNA Interference
Small Interfering RNA
Protein Methyltransferases
Optical Imaging
Gene Silencing
Regulator Genes
Glioblastoma
Blood-Brain Barrier
Luciferases
MicroRNAs
DNA Repair
Veins

Keywords

  • Bioluminescence
  • Glioblastoma multiforme
  • Near-infrared fluorescence
  • O-methylguanine-DNA-methyltransferase
  • Spherical nucleic acid nanoconjugates

ASJC Scopus subject areas

  • General

Cite this

Sita, Timothy L. ; Kouri, Foteini ; Hurley, Lisa A. ; Merkel, Timothy J. ; Chalastanis, Alexandra ; May, Jasmine L. ; Ghelfi, Serena T. ; Cole, Lisa E. ; Cayton, Thomas C. ; Barnaby, Stacey N. ; Sprangers, Anthony J. ; Savalia, Nikunjkumar ; James, Charles David ; Lee, Andrew ; Mirkin, Chad A ; Stegh, Alexander H. / Dual bioluminescence and near-infrared fluorescence monitoring to evaluate spherical nucleic acid nanoconjugate activity in vivo. In: Proceedings of the National Academy of Sciences of the United States of America. 2017 ; Vol. 114, No. 16. pp. 4129-4134.
@article{44018e1b77c54751922f6d1e4c4e1e80,
title = "Dual bioluminescence and near-infrared fluorescence monitoring to evaluate spherical nucleic acid nanoconjugate activity in vivo",
abstract = "RNA interference (RNAi)-based gene regulation platforms have shown promise as a novel class of therapeutics for the precision treatment of cancer. Techniques in preclinical evaluation of RNAibased nanoconjugates have yet to allow for optimization of their gene regulatory activity.We have developed spherical nucleic acids (SNAs) as a blood-brain barrier-/blood-tumor barrier-penetrating nanoconjugate to deliver small interfering (si) and micro (mi)RNAs to intracranial glioblastoma (GBM) tumor sites. To identify high-activity SNA conjugates and to determine optimal SNA treatment regimens, we developed a reporter xenograft model to evaluate SNA efficacy in vivo. Engrafted tumors stably coexpress optical reporters for luciferase and a nearinfrared (NIR) fluorescent protein (iRFP670), with the latter fused to the DNA repair protein O6-methylguanine-DNA-methyltransferase (MGMT). Using noninvasive imaging of animal subjects bearing reporter-modified intracranial xenografts, we quantitatively assessed MGMT knockdown by SNAs composed of MGMT-targeting siRNA duplexes (siMGMT-SNAs). We show that systemic administration of siMGMT-SNAs via single tail vein injection is capable of robust intratumoral MGMT protein knockdown in vivo, with persistent and SNA dose-dependent MGMT silencing confirmed by Western blotting of tumor tissue ex vivo. Analyses of SNA biodistribution and pharmacokinetics revealed rapid intratumoral uptake and significant intratumoral retention that increased the antitumor activity of coadministered temozolomide (TMZ). Our study demonstrates that dual noninvasive bioluminescence and NIR fluorescence imaging of cancer xenograft models represents a powerful in vivo strategy to identify RNAi-based nanotherapeutics with potent gene silencing activity and will inform additional preclinical and clinical investigations of these constructs.",
keywords = "Bioluminescence, Glioblastoma multiforme, Near-infrared fluorescence, O-methylguanine-DNA-methyltransferase, Spherical nucleic acid nanoconjugates",
author = "Sita, {Timothy L.} and Foteini Kouri and Hurley, {Lisa A.} and Merkel, {Timothy J.} and Alexandra Chalastanis and May, {Jasmine L.} and Ghelfi, {Serena T.} and Cole, {Lisa E.} and Cayton, {Thomas C.} and Barnaby, {Stacey N.} and Sprangers, {Anthony J.} and Nikunjkumar Savalia and James, {Charles David} and Andrew Lee and Mirkin, {Chad A} and Stegh, {Alexander H}",
year = "2017",
month = "4",
day = "18",
doi = "10.1073/pnas.1702736114",
language = "English (US)",
volume = "114",
pages = "4129--4134",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "16",

}

Dual bioluminescence and near-infrared fluorescence monitoring to evaluate spherical nucleic acid nanoconjugate activity in vivo. / Sita, Timothy L.; Kouri, Foteini; Hurley, Lisa A.; Merkel, Timothy J.; Chalastanis, Alexandra; May, Jasmine L.; Ghelfi, Serena T.; Cole, Lisa E.; Cayton, Thomas C.; Barnaby, Stacey N.; Sprangers, Anthony J.; Savalia, Nikunjkumar; James, Charles David; Lee, Andrew; Mirkin, Chad A; Stegh, Alexander H.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 114, No. 16, 18.04.2017, p. 4129-4134.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Dual bioluminescence and near-infrared fluorescence monitoring to evaluate spherical nucleic acid nanoconjugate activity in vivo

AU - Sita, Timothy L.

AU - Kouri, Foteini

AU - Hurley, Lisa A.

AU - Merkel, Timothy J.

AU - Chalastanis, Alexandra

AU - May, Jasmine L.

AU - Ghelfi, Serena T.

AU - Cole, Lisa E.

AU - Cayton, Thomas C.

AU - Barnaby, Stacey N.

AU - Sprangers, Anthony J.

AU - Savalia, Nikunjkumar

AU - James, Charles David

AU - Lee, Andrew

AU - Mirkin, Chad A

AU - Stegh, Alexander H

PY - 2017/4/18

Y1 - 2017/4/18

N2 - RNA interference (RNAi)-based gene regulation platforms have shown promise as a novel class of therapeutics for the precision treatment of cancer. Techniques in preclinical evaluation of RNAibased nanoconjugates have yet to allow for optimization of their gene regulatory activity.We have developed spherical nucleic acids (SNAs) as a blood-brain barrier-/blood-tumor barrier-penetrating nanoconjugate to deliver small interfering (si) and micro (mi)RNAs to intracranial glioblastoma (GBM) tumor sites. To identify high-activity SNA conjugates and to determine optimal SNA treatment regimens, we developed a reporter xenograft model to evaluate SNA efficacy in vivo. Engrafted tumors stably coexpress optical reporters for luciferase and a nearinfrared (NIR) fluorescent protein (iRFP670), with the latter fused to the DNA repair protein O6-methylguanine-DNA-methyltransferase (MGMT). Using noninvasive imaging of animal subjects bearing reporter-modified intracranial xenografts, we quantitatively assessed MGMT knockdown by SNAs composed of MGMT-targeting siRNA duplexes (siMGMT-SNAs). We show that systemic administration of siMGMT-SNAs via single tail vein injection is capable of robust intratumoral MGMT protein knockdown in vivo, with persistent and SNA dose-dependent MGMT silencing confirmed by Western blotting of tumor tissue ex vivo. Analyses of SNA biodistribution and pharmacokinetics revealed rapid intratumoral uptake and significant intratumoral retention that increased the antitumor activity of coadministered temozolomide (TMZ). Our study demonstrates that dual noninvasive bioluminescence and NIR fluorescence imaging of cancer xenograft models represents a powerful in vivo strategy to identify RNAi-based nanotherapeutics with potent gene silencing activity and will inform additional preclinical and clinical investigations of these constructs.

AB - RNA interference (RNAi)-based gene regulation platforms have shown promise as a novel class of therapeutics for the precision treatment of cancer. Techniques in preclinical evaluation of RNAibased nanoconjugates have yet to allow for optimization of their gene regulatory activity.We have developed spherical nucleic acids (SNAs) as a blood-brain barrier-/blood-tumor barrier-penetrating nanoconjugate to deliver small interfering (si) and micro (mi)RNAs to intracranial glioblastoma (GBM) tumor sites. To identify high-activity SNA conjugates and to determine optimal SNA treatment regimens, we developed a reporter xenograft model to evaluate SNA efficacy in vivo. Engrafted tumors stably coexpress optical reporters for luciferase and a nearinfrared (NIR) fluorescent protein (iRFP670), with the latter fused to the DNA repair protein O6-methylguanine-DNA-methyltransferase (MGMT). Using noninvasive imaging of animal subjects bearing reporter-modified intracranial xenografts, we quantitatively assessed MGMT knockdown by SNAs composed of MGMT-targeting siRNA duplexes (siMGMT-SNAs). We show that systemic administration of siMGMT-SNAs via single tail vein injection is capable of robust intratumoral MGMT protein knockdown in vivo, with persistent and SNA dose-dependent MGMT silencing confirmed by Western blotting of tumor tissue ex vivo. Analyses of SNA biodistribution and pharmacokinetics revealed rapid intratumoral uptake and significant intratumoral retention that increased the antitumor activity of coadministered temozolomide (TMZ). Our study demonstrates that dual noninvasive bioluminescence and NIR fluorescence imaging of cancer xenograft models represents a powerful in vivo strategy to identify RNAi-based nanotherapeutics with potent gene silencing activity and will inform additional preclinical and clinical investigations of these constructs.

KW - Bioluminescence

KW - Glioblastoma multiforme

KW - Near-infrared fluorescence

KW - O-methylguanine-DNA-methyltransferase

KW - Spherical nucleic acid nanoconjugates

UR - http://www.scopus.com/inward/record.url?scp=85017620761&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85017620761&partnerID=8YFLogxK

U2 - 10.1073/pnas.1702736114

DO - 10.1073/pnas.1702736114

M3 - Article

VL - 114

SP - 4129

EP - 4134

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 16

ER -