Journal Article
. 2019 Nov; 9(26):8332-8343.
doi: 10.7150/thno.35280.

Nanotopography-based lymphatic delivery for improved anti-tumor responses to checkpoint blockade immunotherapy

Sunkuk Kwon 1 Fred Christian Velasquez 1 John C Rasmussen 1 Matthew R Greives 2 Kelly D Turner 2 John R Morrow 1 Wen-Jen Hwu 3 Russell F Ross 4 Songlin Zhang 5 Eva M Sevick-Muraca 1 
  • PMID: 31754400
  •     39 References
  •     11 citations


Rationale: Cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) is a co-inhibitory checkpoint receptor that is expressed by naïve T-cells in lymph nodes (LNs) to inhibit activation against "self" antigens (Ags). In cancer, anti-CTLA-4 blocks inhibitory action, enabling robust activation of T-cells against tumor Ags presented in tumor draining LNs (TDLNs). However, anti-CTLA-4 is administered intravenously with limited exposure within TDLNs and immune related adverse events (irAEs) are associated with over-stimulation of the immune system. Methods: Herein, we first deliver anti-CTLA-4 in an orthotopic mammary carcinoma murine model using a nanotopographical microneedle-array device to compare its anti-tumor response to that from systemic administration. Additionally, to demonstrate the feasibility of lymphatic delivery in humans using the device, we use near-infrared fluorescence imaging to image delivery of ICG to LNs. Results: Our data show that lymphatic infusion results in more effective tumor growth inhibition, arrest of metastases, increased tumor infiltrating lymphocytes and complete responses when compared to conventional systemic administration. In clinical studies, we demonstrate for the first time that nanotopographic infusion can deliver ICG through the lymphatics directly to the axilla and inguinal LNs of healthy human volunteers. Conclusion: Taken together, these results suggest that regional delivery using a nanotopography-based microneedle array could revolutionize checkpoint blockade immunotherapy by reducing systemic drug exposure and maximizing drug delivery to TDLNs where tumor Ags present. Future work is needed to determine whether lymphatic delivery of anti-CTLA-4 can alleviate irAEs that occur with systemic dosing.

Keywords: Fluorescence imaging; Immunotherapy; Lymphatic delivery; Nanotopography.

Anti-programmed cell death-1 (PD-1) monoclonal antibodies in treating advanced melanoma.
Whitney Metcalfe, Jaime Anderson, Van Anh Trinh, Wen-Jen Hwu.
Discov Med, 2015 Jun 25; 19(106). PMID: 26105702
Translation of near-infrared fluorescence imaging technologies: emerging clinical applications.
E M Sevick-Muraca.
Annu Rev Med, 2011 Nov 01; 63. PMID: 22034868
Highly Cited. Review.
Microvascular fluid exchange and the revised Starling principle.
J Rodney Levick, C Charles Michel.
Cardiovasc Res, 2010 Mar 05; 87(2). PMID: 20200043
Highly Cited. Review.
Estimation of rat mammary tumor volume using caliper and ultrasonography measurements.
Ana Faustino-Rocha, Paula A Oliveira, +8 authors, Mário Ginja.
Lab Anim (NY), 2013 May 22; 42(6). PMID: 23689461
Highly Cited.
Interpretation of visual analog scale ratings and change scores: a reanalysis of two clinical trials of postoperative pain.
Mark P Jensen, Connie Chen, Andrew M Brugger.
J Pain, 2003 Nov 19; 4(7). PMID: 14622683
Highly Cited.
In situ formed reactive oxygen species-responsive scaffold with gemcitabine and checkpoint inhibitor for combination therapy.
Chao Wang, Jinqiang Wang, +9 authors, Zhen Gu.
Sci Transl Med, 2018 Feb 23; 10(429). PMID: 29467299
Highly Cited.
T cell-targeting nanoparticles focus delivery of immunotherapy to improve antitumor immunity.
Daniela Schmid, Chun Gwon Park, +9 authors, Michael S Goldberg.
Nat Commun, 2017 Nov 25; 8(1). PMID: 29170511    Free PMC article.
Highly Cited.
Dual-labeled trastuzumab-based imaging agent for the detection of human epidermal growth factor receptor 2 overexpression in breast cancer.
Lakshmi Sampath, Sunkuk Kwon, +4 authors, Eva M Sevick-Muraca.
J Nucl Med, 2007 Sep 06; 48(9). PMID: 17785729
Lymphatic endothelial cells - key players in regulation of tolerance and immunity.
Eric F Tewalt, Jarish N Cohen, Sherin J Rouhani, Victor H Engelhard.
Front Immunol, 2012 Oct 13; 3. PMID: 23060883    Free PMC article.
Enhanced Cancer Immunotherapy by Microneedle Patch-Assisted Delivery of Anti-PD1 Antibody.
Chao Wang, Yanqi Ye, +2 authors, Zhen Gu.
Nano Lett, 2016 Mar 22; 16(4). PMID: 26999507
Highly Cited.
Liposomes to target the lymphatics by subcutaneous administration.
C Oussoren, G Storm.
Adv Drug Deliv Rev, 2001 Aug 08; 50(1-2). PMID: 11489337
Fc-dependent depletion of tumor-infiltrating regulatory T cells co-defines the efficacy of anti-CTLA-4 therapy against melanoma.
Tyler R Simpson, Fubin Li, +11 authors, Sergio A Quezada.
J Exp Med, 2013 Jul 31; 210(9). PMID: 23897981    Free PMC article.
Highly Cited.
The blockade of immune checkpoints in cancer immunotherapy.
Drew M Pardoll.
Nat Rev Cancer, 2012 Mar 23; 12(4). PMID: 22437870    Free PMC article.
Highly Cited. Review.
Lymph node homing of T cells and dendritic cells via afferent lymphatics.
Reinhold Förster, Asolina Braun, Tim Worbs.
Trends Immunol, 2012 Mar 31; 33(6). PMID: 22459312
Highly Cited. Review.
Lymphatic delivery of etanercept via nanotopography improves response to collagen-induced arthritis.
Melissa B Aldrich, Fred C Velasquez, +8 authors, E M Sevick-Muraca.
Arthritis Res Ther, 2017 Jun 02; 19(1). PMID: 28566090    Free PMC article.
Assessing Immune-Related Adverse Events of Efficacious Combination Immunotherapies in Preclinical Models of Cancer.
Jing Liu, Stephen J Blake, +7 authors, Michele W L Teng.
Cancer Res, 2016 Aug 10; 76(18). PMID: 27503925
Pharmacokinetic modeling of the subcutaneous absorption of therapeutic proteins.
Leonid Kagan.
Drug Metab Dispos, 2014 Aug 15; 42(11). PMID: 25122564
Local CTLA4 blockade effectively restrains experimental pancreatic adenocarcinoma growth in vivo.
Linda C Sandin, Fredrik Eriksson, +3 authors, Sara M Mangsbo.
Oncoimmunology, 2014 Apr 05; 3(1). PMID: 24701377    Free PMC article.
Safety profiles of anti-CTLA-4 and anti-PD-1 antibodies alone and in combination.
Celine Boutros, Ahmad Tarhini, +14 authors, Caroline Robert.
Nat Rev Clin Oncol, 2016 May 05; 13(8). PMID: 27141885
Highly Cited. Review.
Immunotherapeutic uses of CpG oligodeoxynucleotides.
Dennis M Klinman.
Nat Rev Immunol, 2004 Apr 02; 4(4). PMID: 15057783
Highly Cited. Review.
Cost-Effectiveness of Nivolumab-Ipilimumab Combination Therapy Compared with Monotherapy for First-Line Treatment of Metastatic Melanoma in the United States.
Anna Oh, Dang M Tran, +4 authors, Leslie Wilson.
J Manag Care Spec Pharm, 2017 May 23; 23(6). PMID: 28530525    Free PMC article.
Nanotopography facilitates in vivo transdermal delivery of high molecular weight therapeutics through an integrin-dependent mechanism.
Laura Walsh, Jubin Ryu, +4 authors, Tejal Desai.
Nano Lett, 2015 Mar 20; 15(4). PMID: 25790174    Free PMC article.
Combination therapy of established tumors by antibodies targeting immune activating and suppressing molecules.
Kazuyoshi Takeda, Yuko Kojima, +7 authors, Mark J Smyth.
J Immunol, 2010 Apr 20; 184(10). PMID: 20400706
Powerful anti-colon cancer effect of modified nanoparticle-mediated IL-15 immunogene therapy through activation of the host immune system.
Xiaoxiao Liu, Yanyan Li, +10 authors, Yuquan Wei.
Theranostics, 2018 Jul 22; 8(13). PMID: 30026861    Free PMC article.
Eradication of established tumors in mice by a combination antibody-based therapy.
Tomoyasu Uno, Kazuyoshi Takeda, +7 authors, Mark J Smyth.
Nat Med, 2006 May 09; 12(6). PMID: 16680149
Immune related adverse events associated with anti-CTLA-4 antibodies: systematic review and meta-analysis.
Anne Bertrand, Marie Kostine, +2 authors, Thierry Schaeverbeke.
BMC Med, 2015 Sep 05; 13. PMID: 26337719    Free PMC article.
Highly Cited. Systematic Review.
Determining the Performance of Fluorescence Molecular Imaging Devices Using Traceable Working Standards With SI Units of Radiance.
Banghe Zhu, John C Rasmussen, Maritoni Litorja, Eva M Sevick-Muraca.
IEEE Trans Med Imaging, 2015 Nov 10; 35(3). PMID: 26552078    Free PMC article.
Nivolumab and ipilimumab versus ipilimumab in untreated melanoma.
Michael A Postow, Jason Chesney, +18 authors, F Stephen Hodi.
N Engl J Med, 2015 Apr 22; 372(21). PMID: 25891304    Free PMC article.
Highly Cited.
Binimetinib versus dacarbazine in patients with advanced NRAS-mutant melanoma (NEMO): a multicentre, open-label, randomised, phase 3 trial.
Reinhard Dummer, Dirk Schadendorf, +21 authors, Keith Flaherty.
Lancet Oncol, 2017 Mar 13; 18(4). PMID: 28284557
Highly Cited.
Eradication of metastatic mouse cancers resistant to immune checkpoint blockade by suppression of myeloid-derived cells.
KiBem Kim, Andrew D Skora, +8 authors, Shibin Zhou.
Proc Natl Acad Sci U S A, 2014 Jul 30; 111(32). PMID: 25071169    Free PMC article.
Highly Cited.
Combined Nivolumab and Ipilimumab or Monotherapy in Untreated Melanoma.
James Larkin, F Stephen Hodi, Jedd D Wolchok.
N Engl J Med, 2015 Sep 24; 373(13). PMID: 26398076
Highly Cited.
Glycosylation and stabilization of programmed death ligand-1 suppresses T-cell activity.
Chia-Wei Li, Seung-Oe Lim, +21 authors, Mien-Chie Hung.
Nat Commun, 2016 Aug 31; 7. PMID: 27572267    Free PMC article.
Highly Cited.
Pembrolizumab versus Ipilimumab in Advanced Melanoma.
Caroline Robert, Jacob Schachter, +20 authors, KEYNOTE-006 investigators.
N Engl J Med, 2015 Apr 22; 372(26). PMID: 25891173
Highly Cited.
The lymphatic system in body homeostasis: physiological conditions.
Waldemar L Olszewski.
Lymphat Res Biol, 2004 Dec 31; 1(1). PMID: 15624317
Emerging lymphatic imaging technologies for mouse and man.
Eva M Sevick-Muraca, Sunkuk Kwon, John C Rasmussen.
J Clin Invest, 2014 Mar 05; 124(3). PMID: 24590275    Free PMC article.
Powerful Anticolon Tumor Effect of Targeted Gene Immunotherapy Using Folate-Modified Nanoparticle Delivery of CCL19 To Activate the Immune System.
Xiaoxiao Liu, Bilan Wang, +7 authors, Yuquan Wei.
ACS Cent Sci, 2019 Mar 06; 5(2). PMID: 30834316    Free PMC article.
Tumor-infiltrating immune cells as potential biomarkers predicting response to treatment and survival in patients with metastatic melanoma receiving ipilimumab therapy.
Tímea Balatoni, Anita Mohos, +8 authors, Andrea Ladányi.
Cancer Immunol Immunother, 2017 Oct 11; 67(1). PMID: 28988380
Tumor-draining lymph nodes are pivotal in PD-1/PD-L1 checkpoint therapy.
Marieke F Fransen, Mark Schoonderwoerd, +5 authors, Ferry Ossendorp.
JCI Insight, 2018 Dec 07; 3(23). PMID: 30518694    Free PMC article.
Highly Cited.
Assessment of lymphatic contractile function after manual lymphatic drainage using near-infrared fluorescence imaging.
I-Chih Tan, Erik A Maus, +6 authors, Eva M Sevick-Muraca.
Arch Phys Med Rehabil, 2011 May 03; 92(5). PMID: 21530723    Free PMC article.
Ruffles and spikes: Control of tight junction morphology and permeability by claudins.
K Sabrina Lynn, Raven J Peterson, Michael Koval.
Biochim Biophys Acta Biomembr, 2020 May 12; 1862(9). PMID: 32389670    Free PMC article.
Dissolving Microneedles with Spatiotemporally controlled pulsatile release Nanosystem for Synergistic Chemo-photothermal Therapy of Melanoma.
Wanbing Qin, Guilan Quan, +7 authors, Chuanbin Wu.
Theranostics, 2020 Jul 30; 10(18). PMID: 32724465    Free PMC article.
The Development and Treatment of Lymphatic Dysfunction in Cancer Patients and Survivors.
Melissa B Aldrich, John C Rasmussen, +2 authors, Eva M Sevick-Muraca.
Cancers (Basel), 2020 Aug 23; 12(8). PMID: 32823928    Free PMC article.
Nano-Enhanced Cancer Immunotherapy: Immunology Encounters Nanotechnology.
Ernesto Bockamp, Sebastian Rosigkeit, Dominik Siegl, Detlef Schuppan.
Cells, 2020 Sep 19; 9(9). PMID: 32942725    Free PMC article.
Cell and tissue engineering in lymph nodes for cancer immunotherapy.
Alexander J Najibi, David J Mooney.
Adv Drug Deliv Rev, 2020 Aug 05; 161-162. PMID: 32750376    Free PMC article.
Systematic Review.
Reducing PD-L1 expression with a self-assembled nanodrug: an alternative to PD-L1 antibody for enhanced chemo-immunotherapy.
Shuxian Cai, Ziyi Chen, +6 authors, Huanghao Yang.
Theranostics, 2021 Jan 08; 11(4). PMID: 33408792    Free PMC article.
New Therapeutic Options for Advanced Hepatocellular Carcinoma.
Yu-Shui Ma, Ji-Bin Liu, Ting-Miao Wu, Da Fu.
Cancer Control, 2020 Aug 18; 27(3). PMID: 32799550    Free PMC article.
The future of microfluidics in immune checkpoint blockade.
Jonathan Briones, Wilfred Espulgar, +3 authors, Masato Saito.
Cancer Gene Ther, 2020 Oct 29; 28(9). PMID: 33110208
Above the Matrix: Functional Roles for Apically Localized Integrins.
Raven J Peterson, Michael Koval.
Front Cell Dev Biol, 2021 Sep 07; 9. PMID: 34485286    Free PMC article.
Microneedle arrays integrated with living organisms for smart biomedical applications.
Bo Cai, Yusheng Gong, +2 authors, Wei Chen.
Theranostics, 2021 Nov 25; 11(20). PMID: 34815801    Free PMC article.
Enhanced T-Cell Priming and Improved Anti-Tumor Immunity through Lymphatic Delivery of Checkpoint Blockade Immunotherapy.
Carolina Mantilla-Rojas, Fred C Velasquez, +4 authors, Eva M Sevick-Muraca.
Cancers (Basel), 2022 Apr 13; 14(7). PMID: 35406595    Free PMC article.