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Membrane-bound MMP-14 protease-activatable adeno-associated viral vectors for gene delivery to pancreatic tumors

Gene Therapy volume 29, pages 138–146 (2022)Cite this article

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Abstract

Adeno-associated virus’ (AAV) relatively simple structure makes it accommodating for engineering into controllable delivery platforms. Cancer, such as pancreatic ductal adenocarcinoma (PDAC), are often characterized by upregulation of membrane-bound proteins, such as MMP-14, that propagate survival integrin signaling. In order to target tumors, we have engineered an MMP-14 protease-activatable AAV vector that responds to both membrane-bound and extracellularly active MMPs. This “provector” was generated by inserting a tetra-aspartic acid inactivating motif flanked by the MMP-14 cleavage sequence IPESLRAG into the capsid subunits. The MMP-14 provector shows lower background transduction than previously developed provectors, leading to a 9.5-fold increase in transduction ability. In a murine model of PDAC, the MMP-14 provector shows increased delivery to an allograft tumor. This proof-of-concept study illustrates the possibilities of membrane-bound protease-activatable gene therapies to target tumors.

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Fig. 1: Provector design and production characterization.
Fig. 2: In vitro performance of IPES provector.
Fig. 3: Development of negative control vector with scrambled protease cleavage sequence.
Fig. 4: In vivo performance of provectors in murine model of PDAC.
Fig. 5: Non-tumor mouse in vivo performance of provector.

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References

  1. Samulski R, Muzyczka N. AAV-mediated gene therapy for research and therapeutic purposes. Annu Rev Virol. 2014;1:427–51.

    Article  PubMed  Google Scholar 

  2. Bi L. Summary basis for regulatory action -Luxturna. Food and Drug Administration (FDA), May, 2019.

  3. Byrnes A. Summary basis for regulatory action - Zolgensma. Food and Drug Administration (FDA), May, 2019.

  4. Xiong W, Wu D, Xue Y, Wang S, Chung M, Ji X, et al. AAV Cis-regulatory sequences are correlated with ocular toxicity. Proc Natl Acad Sci USA. 2019;116:5785–94.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Evans A, Thadani N, Suh J. Biocomputing nanoplatforms as therapeutics and diagnostics.J. Control. Release Soc.2016;240:387–93.

    Article  CAS  Google Scholar 

  6. Siegel R, Miller K, Jemal A. Cancer statistics, 2020. CA. Cancer J Clin. 2020;70:7–30.

    Article  PubMed  Google Scholar 

  7. Tian C, Öhlund D, Rickelt S, Lidström T, Huang Y, Hao L, et al. Cancer-cell-derived matrisome proteins promote metastasis in pancreatic ductal adenocarcinoma. Cancer Res. 2020;80:1461–147.

  8. Cid-Arregui A, Juarez V. Perspectives in the treatment of pancreatic adenocarcinoma. World J Gastroenterol. 2015;21:9297–316.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Ellenrieder V, Alber B, Lacher U, Hendler S, Menke A, Boeck W, et al. Role of MT-MMPs and MMP-2 in pancreatic cancer progression. Int J Cancer. 2000;85:14–20.

  10. Jiang W, Zhang Y, Kane K, Collins M, Simeone D, Di Magliano M, et al. CD44 regulates pancreatic cancer invasion through MT1-MMP. Mol Cancer Res. 2015;13:9–15.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Cepeda M, Pelling J, Evered C, Williams K, Freedman Z, Stan I, et al. Less is more: low expression of MT1-MMP is optimal to promote migration and tumourigenesis of breast cancer cells. Mol Cancer. 2016;15.

  12. Vihinen P, Kähäri V. Matrix metalloproteinases in cancer: prognostic markers and therapeutic targets. Int J Cancer. 2002;99:157–66.

  13. Guenther C, Brun M, Bennett A, Ho M, Chen W, Zhu B, et al. Protease-activatable adeno-associated virus vector for gene delivery to damaged heart tissue. Mol Ther. 2019;27:611–22.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Chen M, Robinson T, Suh J. Longer inactivating sequence in peptide lock improves performance of synthetic protease-activatable adeno-associated virus. ACS Synth Biol. 2019;8:91–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Robinson T, Judd J, Ho M, Suh J. Role of tetra amino acid motif properties on the function of protease-activatable viral vectors. ACS Biomater Sci Eng. 2016;2:2026–33.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Judd J, Ho M, Tiwari A, Gomez E, Dempsey C, Van Vliet K, et al. Tunable protease-activatable virus nanonodes. ACS Nano. 2014;8:4740–6.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Ho M, Adler B, Torre M, Silberg J, Suh J. SCHEMA computational design of virus capsid chimeras: calibrating how genome packaging, protection, and transduction correlate with calculated structural disruption. ACS Synth Biol. 2013;2:724–33.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Turk B, Huang L, Piro E, Cantley L. Determination of protease cleavage site motifs using mixture-based oriented peptide libraries. Nat Biotechnol. 2001;19:661.

    Article  CAS  PubMed  Google Scholar 

  19. Li J, Qian W, Qin T, Xiao Y, Cheng L, Cao J, et al. Mouse-derived allografts: a complementary model to the KPC mice on researching pancreatic cancer in vivo. Comput Struct Biotechnol J. 2019;17:498–506.

    Article  PubMed  PubMed Central  Google Scholar 

  20. Erstad D, Sojoodi M, Taylor M, Ghoshal S, Razavi A, Graham-O’Regan K, et al. Orthotopic and heterotopic murine models of pancreatic cancer and their different responses to FOLFIRINOX chemotherapy. Dis Model Mech. 2018;11.

  21. Chen S, Johnston J, Sandhu A, Bish L, HovHannisyan R, Jno-Charles O, et al. Enhancing the utility of adeno-associated virus gene transfer through inducible tissue-specific expression. Hum Gene Ther Methods. 2013;24:270–8.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Pacak C, Sakai Y, Thattaliyath B, Mah C, Byrne B. Tissue specific promoters improve specificity of aav9 mediated transgene expression following intra-vascular gene delivery in neonatal mice. Genet Vaccines Ther. 2008;6.

  23. Krantz S, Shields M, Dangi-Garimella S, Cheon E, Barron M, Hwang R, et al. MT1-MMP cooperates with KrasG12D to promote pancreatic fibrosis through increased TGF-β signaling. Mol Cancer Res. 2011;9:1294–304.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Kozera B, Rapacz M.Reference genes in real-time PCR.J Appl Genet.2013;54:391–406.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Tong J, Evans A, Ho M, Guenther C, Brun M, Judd J, et al. Reducing off target viral delivery in ovarian cancer gene therapy using a protease-activated AAV2 vector platform. J Control Release. 2019;307:292–301.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Kanazawa T, Mizukami H, Okada T, Hanazono Y, Kume A, Nishino H, et al. Suicide gene therapy using AAV-HSVtk/ganciclovir in combination with irradiation results in regression of human head and neck cancer xenografts in nude mice. Gene Ther. 2003;10:51–8.

    Article  CAS  PubMed  Google Scholar 

  27. Reul J, Frisch J, Engeland C, Thalheimer F, Hartmann J, Ungerechts G, et al. Tumor-specific delivery of immune checkpoint inhibitors by engineered AAV vectors. Front Oncol. 2019;9.

  28. Jiang M, Liu Z, Xiang Y, Ma H, Liu S, Liu Y, et al. Synergistic antitumor effect of AAV-mediated TRAIL expression combined with cisplatin on head and neck squamous cell carcinoma. BMC Cancer. 2011;11.

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Acknowledgements

We acknowledge University of North Carolina at Chapel Hill Gene Therapy Center Core for providing us with pXX6-80 and scAAV2-CMV-GFP, the University of Pennsylvania Vector Core for providing us with pAAV2/9, and the University of Iowa Vector Core for providing us with ITR-CMV-mCherry.

Funding

Funding

NIH (R01CA207497) to JS, NCI MD Anderson/Rice Cancer T32 Fellowship 5T32CA196561-05 to SB, and NIH (R01CA220236 and R01CA218403) to A.M.

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Authors and Affiliations

  1. Department of Bioengineering, Rice University, Houston, TX, USA

    Susan S. Butler, Cooper Lueck & Junghae Suh

  2. Departments of Translational Molecular Pathology and Pathology, Sheikh Ahmed Center for Pancreatic Cancer Research, The University of Texas MD Anderson Cancer Center, Houston, TX, USA

    Kenjiro Date, Takashi Okumura, Bidyut Ghosh & Anirban Maitra

  3. Systems, Synthetic, and Physical Biology, Rice University, Houston, TX, USA

    Junghae Suh

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  1. Susan S. Butler
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Correspondence to Junghae Suh.

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Conflict of interest

JS is an employee of Biogen as of August, 2019. AM receives royalties for a pancreatic cancer biomarker test from Cosmos Wisdom Biotechnology, and this financial relationship is managed and monitored by the UTMDACC Conflict of Interest Committee. AM is also listed as an inventor on a patent that has been licensed by Johns Hopkins University to ThriveEarlier Detection.

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Butler, S.S., Date, K., Okumura, T. et al. Membrane-bound MMP-14 protease-activatable adeno-associated viral vectors for gene delivery to pancreatic tumors. Gene Ther 29, 138–146 (2022). https://doi.org/10.1038/s41434-021-00255-9

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