Genetically Engineered Nanoparticles

Our research in this area uses hybrid recombinant peptide polymers, the composition of which are inspired by Elastin-like Polypeptides (ELPs), Resilin-like polypeptide (RLPs), and their fusion with lipids, and bioactive peptides. By tuning the sequence and composition at the gene level we can tune their self-assembly with near absolute precision. Our goal is to find the “magic bullet” for anti-cancer therapy. We envision achieving this in four stages. In stage one we strive to increase passive targeting to the tumor: (i) by precise engineering of self-assembled particles in terms of shape, size, aspect ratio, mobility, dispersity and surface-charge density by exploiting phase transition behavior of the parent peptide/hybrid peptide backbones, and (ii) by incorporating stealth behavior to the nanoparticle by using a long-circulating peptide domain (like albumin-binding domain) or by redesigning the parent peptide. We showed that self-assembly of a zwitterionic peptide polymer that mimics synthetic PEG to significantly increase the plasma half-life of the nanoparticle, potentially avoiding antigenicity. In stage two we pursue homing the nanoparticle to tumor cells by decorating the nanoparticle with a tumor selective peptide-ligand at gene level. This avoids the drawbacks of covalent conjugation of ligand moiety, such as limited control of ligand valency, use of excess of ligand, manufacturing hassle etc. The targeting ability not only depends on ligand affinity but also on valency, shape of the particles and the linker with which it is attached to the particle surface. In stage three we aim to precisely optimize the drug binding domain (using lipids, canonical and non-canonical amino acids, cleavable linker etc) to mask small molecules in the nanoparticle core avoiding their premature leakage. Our motivation to demonstrate the versatility of our technology is reflected in our choice of drug that ranges from hydrophobic paclitaxel to hydrophilic gemcitabine, from bioactive peptide to inorganic metal ion like Pt. An ongoing effort is now directed towards combining this technology with our smart, tunable immunotherapy making it stage four in combating an established tumor.

graphic diagram of 4 stages

Publications

J. Haley, Jones, J. B. , Petraki, S. , Callander, M. , Shrestha, S. , Springfield, E. , Adamson, L. , Chilkoti, A. , Dzuricky, M. J. , and Luginbuhl, K. M. , “IsoTag™AAV: an innovative, scalable & non-chromatographic method for streamlined AAV manufacturing”, Cell & Gene Therapy Insights, vol. 8, no. 10, pp. 1287-1300, 2022.

Schaal, Jeffrey L., Jayanta Bhattacharyya, Jeremy Brownstein, Kyle C. Strickland, Garrett Kelly, Soumen Saha, Joshua Milligan, et al. “Brachytherapy via a depot of biopolymer-bound 131I synergizes with nanoparticle paclitaxel in therapy-resistant pancreatic tumours.” Nat Biomed Eng 6, no. 10 (October 2022): 1148–66. https://doi.org/10.1038/s41551-022-00949-4.

Saha, Soumen, Samagya Banskota, Jianqiao Liu, Nikita Zakharov, Michael Dzuricky, Xinghai Li, Ping Fan, et al. “Genetically Engineered Nanoparticles of Asymmetric Triblock Polypeptide with a Platinum(IV) Cargo Outperforms a Platinum(II) Analog and Free Drug in a Murine Cancer Model.” Nano Lett 22, no. 14 (July 27, 2022): 5898–5908. https://doi.org/10.1021/acs.nanolett.2c01850.

Weber, Patrick, Michael Dzuricky, Junseon Min, Irene Jenkins, and Ashutosh Chilkoti. “Concentration-Independent Multivalent Targeting of Cancer Cells by Genetically Encoded Core-Crosslinked Elastin/Resilin-like Polypeptide Micelles.” Biomacromolecules 22, no. 10 (October 2021): 4347–56. https://doi.org/10.1021/acs.biomac.1c00897.

Navarro, Luis A., Justin J. Ryan, Michael Dzuricky, Michael Gradzielski, Ashutosh Chilkoti, and Stefan Zauscher. “Microphase Separation of Resilin-like and Elastin-like Diblock Copolypeptides in Concentrated Solutions.” Biomacromolecules 22, no. 9 (September 2021): 3827–38. https://doi.org/10.1021/acs.biomac.1c00672.

Jenkins, Irene C., Joshua J. Milligan, and Ashutosh Chilkoti. “Genetically Encoded Elastin-Like Polypeptides for Drug Delivery.” Advanced Healthcare Materials 10, no. 13 (July 2021): e2100209. https://doi.org/10.1002/adhm.202100209.

Wang, Jing, Junseon Min, Seyed Ali Eghtesadi, Ravi S. Kane, and Ashutosh Chilkoti. “Quantitative Study of the Interaction of Multivalent Ligand-Modified Nanoparticles with Breast Cancer Cells with Tunable Receptor Density.” ACS Nano 14, no. 1 (January 2020): 372–83. https://doi.org/10.1021/acsnano.9b05689.

Wang, Jing, Soumen Saha, Jeffrey L. Schaal, Parisa Yousefpour, Xinghai Li, and Ashutosh Chilkoti. “Heuristics for the Optimal Presentation of Bioactive Peptides on Polypeptide Micelles.” Nano Letters 19, no. 11 (November 2019): 7977–87. https://doi.org/10.1021/acs.nanolett.9b03141.

Manzari, Mandana T., Gray R. Anderson, Kevin H. Lin, Ryan S. Soderquist, Merve Çakir, Mitchell Zhang, Chandler E. Moore, et al. “Genomically informed small-molecule drugs overcome resistance to a sustained-release formulation of an engineered death receptor agonist in patient-derived tumor models.” Sci Adv 5, no. 9 (September 2019): eaaw9162. https://doi.org/10.1126/sciadv.aaw9162.

Dzuricky, Michael, Sinan Xiong, Patrick Weber, and Ashutosh Chilkoti. “Avidity and Cell Uptake of Integrin-Targeting Polypeptide Micelles is Strongly Shape-Dependent.” Nano Letters 19, no. 9 (September 2019): 6124–32. https://doi.org/10.1021/acs.nanolett.9b02095.

Yousefpour, Parisa, Lucie Ahn, Joel Tewksbury, Soumen Saha, Simone A. Costa, Joseph J. Bellucci, Xinghai Li, and Ashutosh Chilkoti. “Conjugate of Doxorubicin to Albumin-Binding Peptide Outperforms Aldoxorubicin.” Small (Weinheim an Der Bergstrasse, Germany) 15, no. 12 (March 2019): e1804452. https://doi.org/10.1002/smll.201804452.

Banskota, Samagya, Parisa Yousefpour, Nadia Kirmani, Xinghai Li, and Ashutosh Chilkoti. “Long circulating genetically encoded intrinsically disordered zwitterionic polypeptides for drug delivery.” Biomaterials 192 (February 2019): 475–85. https://doi.org/10.1016/j.biomaterials.2018.11.012.

Costa, Simone A., Davoud Mozhdehi, Michael J. Dzuricky, Farren J. Isaacs, Eric M. Brustad, and Ashutosh Chilkoti. “Active Targeting of Cancer Cells by Nanobody Decorated Polypeptide Micelle with Bio-orthogonally Conjugated Drug.” Nano Letters 19, no. 1 (January 2019): 247–54. https://doi.org/10.1021/acs.nanolett.8b03837.

Mozhdehi, Davoud, Kelli M. Luginbuhl, Michael Dzuricky, Simone A. Costa, Sinan Xiong, Fred C. Huang, Mae M. Lewis, Stephanie R. Zelenetz, Christian D. Colby, and Ashutosh Chilkoti. “Genetically Encoded Cholesterol-Modified Polypeptides.” Journal of the American Chemical Society 141, no. 2 (January 2019): 945–51. https://doi.org/10.1021/jacs.8b10687.

Yousefpour, Parisa, Jonathan R. McDaniel, Varun Prasad, Lucie Ahn, Xinghai Li, Rishi Subrahmanyan, Isaac Weitzhandler, Steven Suter, and Ashutosh Chilkoti. “Genetically Encoding Albumin Binding into Chemotherapeutic-loaded Polypeptide Nanoparticles Enhances Their Antitumor Efficacy.” Nano Letters 18, no. 12 (December 2018): 7784–93. https://doi.org/10.1021/acs.nanolett.8b03558.

Mozhdehi, Davoud, Kelli M. Luginbuhl, Joseph R. Simon, Michael Dzuricky, Rüdiger Berger, H Samet Varol, Fred C. Huang, et al. “Genetically encoded lipid-polypeptide hybrid biomaterials that exhibit temperature-triggered hierarchical self-assembly.” Nature Chemistry 10, no. 5 (May 2018): 496–505. https://doi.org/10.1038/s41557-018-0005-z.

Costa, Simone A., Joseph R. Simon, Miriam Amiram, Lei Tang, Stefan Zauscher, Eric M. Brustad, Farren J. Isaacs, and Ashutosh Chilkoti. “Photo-Crosslinkable Unnatural Amino Acids Enable Facile Synthesis of Thermoresponsive Nano- to Microgels of Intrinsically Disordered Polypeptides.” Advanced Materials (Deerfield Beach, Fla.) 30, no. 5 (February 2018). https://doi.org/10.1002/adma.201704878.

Luginbuhl, Kelli M., Davoud Mozhdehi, Michael Dzuricky, Parisa Yousefpour, Fred C. Huang, Nicholas R. Mayne, Kristen L. Buehne, and Ashutosh Chilkoti. “Recombinant Synthesis of Hybrid Lipid-Peptide Polymer Fusions that Self-Assemble and Encapsulate Hydrophobic Drugs.” Angewandte Chemie (International Ed. in English) 56, no. 45 (November 2017): 13979–84. https://doi.org/10.1002/anie.201704625.

Wang, Jing, Michael Dzuricky, and Ashutosh Chilkoti. “The Weak Link: Optimization of the Ligand-Nanoparticle Interface To Enhance Cancer Cell Targeting by Polymer Micelles.” Nano Letters 17, no. 10 (October 2017): 5995–6005. https://doi.org/10.1021/acs.nanolett.7b02225.

Bhattacharyya, Jayanta, Isaac Weitzhandler, Shihan Bryan Ho, Jonathan R. McDaniel, Xinghai Li, Lei Tang, Jinyao Liu, Mark Dewhirst, and Ashutosh Chilkoti. “Encapsulating a Hydrophilic Chemotherapeutic into Rod-like Nanoparticles of a Genetically Encoded Asymmetric Triblock Polypeptide Improves its Efficacy.” Adv Funct Mater 27, no. 12 (March 24, 2017). https://doi.org/10.1002/adfm.201605421.

Bhattacharyya, Jayanta, Joseph J. Bellucci, Isaac Weitzhandler, Jonathan R. McDaniel, Ivan Spasojevic, Xinghai Li, Chao-Chieh Lin, Jen-Tsan Ashley Chi, and Ashutosh Chilkoti. “A paclitaxel-loaded recombinant polypeptide nanoparticle outperforms Abraxane in multiple murine cancer models.” Nat Commun 6 (August 4, 2015): 7939. https://doi.org/10.1038/ncomms8939.

Patents