Abstract
Noncompressible torso hemorrhage accounts for a significant portion of preventable trauma deaths. We report here on the development of injectable, targeted supramolecular nanotherapeutics based on peptide amphiphile (PA) molecules that are designed to target tissue factor (TF) and, therefore, selectively localize to sites of injury to slow hemorrhage. Eight TF-targeting sequences were identified, synthesized into PA molecules, coassembled with nontargeted backbone PA at various weight percentages, and characterized via circular dichroism spectroscopy, transmission electron microscopy, and X-ray scattering. Following intravenous injection in a rat liver hemorrhage model, two of these PA nanofiber coassemblies exhibited the most specific localization to the site of injury compared to controls (p < 0.05), as quantified using immunofluorescence imaging of injured liver and uninjured organs. To determine if the nanofibers were targeting TF in vivo, a mouse saphenous vein laser injury model was performed and showed that TF-targeted nanofibers colocalized with fibrin, demonstrating increased levels of nanofiber at TF-rich sites. Thromboelastograms obtained using samples of heparinized rat whole blood containing TF demonstrated that no clots were formed in the absence of TF-targeted nanofibers. Lastly, both PA nanofiber coassemblies decreased blood loss in comparison to sham and backbone nanofiber controls by 35-59% (p < 0.05). These data demonstrate an optimal TF-targeted nanofiber that localizes selectively to sites of injury and TF exposure, and, interestingly, reduces blood loss. This research represents a promising initial phase in the development of a TF-targeted injectable therapeutic to reduce preventable deaths from hemorrhage.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 6649-6662 |
| Number of pages | 14 |
| Journal | ACS nano |
| Volume | 14 |
| Issue number | 6 |
| DOIs | |
| State | Published - Jun 23 2020 |
Funding
This study was supported, in part, by funding from the Department of Defense AFRL/RQKHC (FA8650-16-2-6G19 to B.G., M.R. Kibbe, T.A.P., and S.I.S.), the National Institutes of Health (1R01HL116577-01 to M.R. Kibbe and S.I.S.; 1R35HL144976-01 to W.B.), and the University of North Carolina School of Medicine. M.K.K. and R.H.L. received funding through the UNC Hematology NIH T32 Training Grant (HL007149-42). E.B.P. was supported by the American Heart Association Postdoctoral Fellowship Award 18POST33960499. S.I.S. acknowledges funding from the Louis A. Simpson and Kimberly K. Querrey Center for Regenerative Nanomedicine at Northwestern University through a CRN Catalyst Award. The UNC Electron Microscopy Facility is supported in part by the Lineberger Comprehensive Cancer Center UCRF. Peptide amphiphile synthesis was performed in the Peptide Synthesis Core Facility of the Simpson Querrey Institute at Northwestern University. The U.S. Army Research Office, the U.S. Army Medical Research and Materiel Command, and Northwestern University provided funding to develop this facility, and ongoing support is being received from the Soft and Hybrid Nanotechnology Experimental (SHyNE) Resource (NSF ECCS-1542205). The authors would like to thank K. Wallace and D. Hepp for their administrative assistance with this manuscript. We would also like to thank the UNC Structural Biology Core for their assistance with the SBC-X PA nanofiber sequences. We gratefully acknowledge J. Griffith and S. Willcox in the Lineberger Cancer Center EM Core Facility for assistance with conventional TEM. T.D.C. graciously acknowledges support from an American Australian Association-Dow Chemical Company Scholarship. We acknowledge the following core facilities at Northwestern University: the Peptide Synthesis Core Facility and the Analytical BioNanoTechnology Equipment Core Facility (ANTEC) of the Simpson Querrey Institute for peptide synthesis and purification, and the Biological Imaging Facility (BIF) for cryo-TEM. X-ray scattering experiments were performed at the DuPont–Northwestern–Dow Collaborative Access Team (DND-CAT) located at Sector 5 of the Advanced Photon Source. DND-CAT is supported by E.I. DuPont de Nemours & Co., the Dow Chemical Company, and the State of Illinois. This research used resources of the Advanced Photon Source, a U.S. Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory under Contract No. DE-AC02-06CH11357. Cryo-TEM images made use of the BioCryo facility of Northwestern University’s NUANCE Center, which has received support from the Soft and Hybrid Nanotechnology Experimental (SHyNE) Resource (NSF ECCS-1542205); the MRSEC program (NSFDMR-1720139) at the Materials Research Center; the International Institute for Nanotechnology (IIN); and the State of Illinois, through the IIN. The authors thank N.D. Riggsbee and T.C. Nichols for help and advice with thromboelastography. The views expressed in this article are those of the authors and do not necessarily reflect the official policy or position of the Air Force, the Department of Defense, or the U.S. Government; cleared, SAF/PA, Case #2019-0302, 24 Apr 2019.
Keywords
- battlefield intervention
- hemorrhage
- nanofibers
- noncompressible torso hemorrhage
- preventable deaths
- targeted therapeutic
- tissue factor
ASJC Scopus subject areas
- General Materials Science
- General Engineering
- General Physics and Astronomy
