Conformational ligand-directed targeting of calcium-dependent receptors in acute trauma

Renata Pasqualini¹, Christopher Markosian², Daniela I Staquicini², Andrey S Dobroff³, Esteban Dodero-Rojas⁴, Paul C Whitford⁵, E Magda Barbu³, Julianna K Bronk⁶, Marina Cardó-Vila⁷, Dawn R Christianson³, Emmanuel Dias-Neto⁸, Wouter H P Driessen³, Liliana Guzman-Rojas⁹, Serena Marchiò¹⁰, Diana N Nunes⁸, Francislon S de Oliveira¹¹, Michael G Ozawa¹², Bettina Proneth¹³, Roberto Rangel¹⁴, Tracey L Smith², Glauco R Souza³, Fernanda I Staquicini², Fenny H F Tang², Wallace B Baze¹⁵, João C Setubal¹¹, John W Burns¹⁶, Michael A Dubick¹⁶, Juri G Gelovani¹⁷, Andriy I Batchinsky¹⁸, Jon E Mogford¹⁹, Charles E Wade²⁰, John B Holcomb²¹, Stephen K Burley²², José N Onuchic²³, Wadih Arap²⁴

Affiliations

¹ Rutgers Cancer Institute, Newark, NJ, USA; Division of Cancer Biology, Department of Radiation Oncology, Rutgers New Jersey Medical School, Newark, NJ, USA. Electronic address: renata.pasqualini@rutgers.edu.
² Rutgers Cancer Institute, Newark, NJ, USA; Division of Cancer Biology, Department of Radiation Oncology, Rutgers New Jersey Medical School, Newark, NJ, USA.
³ David H. Koch Center, Houston, TX, USA; Department of Genitourinary Medical Oncology, University of Texas MD Anderson Cancer Center, Houston, TX, USA.
⁴ Center for Theoretical Biological Physics, Rice University, Houston, TX, USA.
⁵ Center for Theoretical Biological Physics, Northeastern University, Boston, MA, USA; Department of Physics, Northeastern University, Boston, MA, USA.
⁶ Department of Radiation Oncology, University of Texas MD Anderson Cancer Center, Houston, TX, USA.
⁷ Department of Cellular & Molecular Medicine, University of Arizona Cancer Center, University of Arizona, Tucson, AZ, USA; Department of Otolaryngology and Head & Neck Surgery, University of Arizona Cancer Center, University of Arizona, Tucson, AZ, USA.
⁸ Rutgers Cancer Institute, Newark, NJ, USA; Division of Cancer Biology, Department of Radiation Oncology, Rutgers New Jersey Medical School, Newark, NJ, USA; Laboratory of Medical Genomics, A.C. Camargo Cancer Center, São Paulo, SP, Brazil.
⁹ Cancer Research Program, Houston Methodist Research Institute, Houston, TX, USA.
¹⁰ Department of Oncology, University of Turin, Candiolo, TO, Italy.
¹¹ Department of Biochemistry, Institute of Chemistry, University of São Paulo, São Paulo, SP, Brazil.
¹² Department of Pathology, Stanford University School of Medicine, Stanford, CA, USA.
¹³ Institute of Metabolism and Cell Death, Helmholtz Center Munich, Neuherberg, BY, Germany.
¹⁴ Department of Head & Neck Surgery, University of Texas MD Anderson Cancer Center, Houston, TX, USA.
¹⁵ Keeling Center for Comparative Medicine & Research, Department of Comparative Medicine, University of Texas MD Anderson Cancer Center, Bastrop, TX, USA.
¹⁶ US Army Institute of Surgical Research, Fort Sam Houston, TX, USA.
¹⁷ Office of the Provost, United Arab Emirates University, Al Ain, Abu Dhabi, UAE; Department of Radiology, College of Medicine & Health Sciences, United Arab Emirates University, Al Ain, Abu Dhabi, UAE; Department of Radiology, Siriraj Hospital, Mahidol University, Bangkok, Thailand; Department of Biomedical Engineering, College of Engineering and School of Medicine, Wayne State University, Detroit, MI, USA.
¹⁸ Autonomous Reanimation & Evacuation (AREVA) Research Institute and Innovation Center, The Geneva Foundation, San Antonio, TX, USA.
¹⁹ Defense Sciences Office, Defense Advanced Research Projects Agency, Arlington, VA, USA.
²⁰ Center for Translational Injury Research, Department of Surgery, McGovern Medical School at the University of Texas Health Science Center, Houston, TX, USA; CAG Center for Endotheliomics, Copenhagen University Hospital - Rigshospitalet, Copenhagen, Denmark; Department of Clinical Medicine, University of Copenhagen, Copenhagen, Denmark.
²¹ Center for Injury Science, University of Alabama at Birmingham, Birmingham, AL, USA; Division of Acute Care Surgery, Department of Surgery, University of Alabama at Birmingham, Birmingham, AL, USA.
²² RCSB Protein Data Bank, Rutgers, The State University of New Jersey, Piscataway, NJ, USA; RCSB Protein Data Bank, San Diego Supercomputer Center, University of California, San Diego, La Jolla, CA, USA; Institute for Quantitative Biomedicine, Rutgers, The State University of New Jersey, Piscataway, NJ, USA; Rutgers Cancer Institute, New Brunswick, NJ, USA.
²³ Center for Theoretical Biological Physics, Rice University, Houston, TX, USA; Department of Biosciences, Rice University, Houston, TX, USA; Department of Chemistry, Rice University, Houston, TX, USA; Department of Physics & Astronomy, Rice University, Houston, TX, USA.
²⁴ Rutgers Cancer Institute, Newark, NJ, USA; Division of Hematology/Oncology, Department of Medicine, Rutgers New Jersey Medical School, Newark, NJ, USA. Electronic address: wadih.arap@rutgers.edu.

PMID: 40609540
DOI: 10.1016/j.medj.2025.100638

Conformational ligand-directed targeting of calcium-dependent receptors in acute trauma

Renata Pasqualini et al. Med. 2025.

. 2025 Jul 11;6(7):100638.

doi: 10.1016/j.medj.2025.100638. Epub 2025 Jul 2.

Authors

Affiliations

¹ Rutgers Cancer Institute, Newark, NJ, USA; Division of Cancer Biology, Department of Radiation Oncology, Rutgers New Jersey Medical School, Newark, NJ, USA. Electronic address: renata.pasqualini@rutgers.edu.
² Rutgers Cancer Institute, Newark, NJ, USA; Division of Cancer Biology, Department of Radiation Oncology, Rutgers New Jersey Medical School, Newark, NJ, USA.
³ David H. Koch Center, Houston, TX, USA; Department of Genitourinary Medical Oncology, University of Texas MD Anderson Cancer Center, Houston, TX, USA.
⁴ Center for Theoretical Biological Physics, Rice University, Houston, TX, USA.
⁵ Center for Theoretical Biological Physics, Northeastern University, Boston, MA, USA; Department of Physics, Northeastern University, Boston, MA, USA.
⁶ Department of Radiation Oncology, University of Texas MD Anderson Cancer Center, Houston, TX, USA.
⁷ Department of Cellular & Molecular Medicine, University of Arizona Cancer Center, University of Arizona, Tucson, AZ, USA; Department of Otolaryngology and Head & Neck Surgery, University of Arizona Cancer Center, University of Arizona, Tucson, AZ, USA.
⁸ Rutgers Cancer Institute, Newark, NJ, USA; Division of Cancer Biology, Department of Radiation Oncology, Rutgers New Jersey Medical School, Newark, NJ, USA; Laboratory of Medical Genomics, A.C. Camargo Cancer Center, São Paulo, SP, Brazil.
⁹ Cancer Research Program, Houston Methodist Research Institute, Houston, TX, USA.
¹⁰ Department of Oncology, University of Turin, Candiolo, TO, Italy.
¹¹ Department of Biochemistry, Institute of Chemistry, University of São Paulo, São Paulo, SP, Brazil.
¹² Department of Pathology, Stanford University School of Medicine, Stanford, CA, USA.
¹³ Institute of Metabolism and Cell Death, Helmholtz Center Munich, Neuherberg, BY, Germany.
¹⁴ Department of Head & Neck Surgery, University of Texas MD Anderson Cancer Center, Houston, TX, USA.
¹⁵ Keeling Center for Comparative Medicine & Research, Department of Comparative Medicine, University of Texas MD Anderson Cancer Center, Bastrop, TX, USA.
¹⁶ US Army Institute of Surgical Research, Fort Sam Houston, TX, USA.
¹⁷ Office of the Provost, United Arab Emirates University, Al Ain, Abu Dhabi, UAE; Department of Radiology, College of Medicine & Health Sciences, United Arab Emirates University, Al Ain, Abu Dhabi, UAE; Department of Radiology, Siriraj Hospital, Mahidol University, Bangkok, Thailand; Department of Biomedical Engineering, College of Engineering and School of Medicine, Wayne State University, Detroit, MI, USA.
¹⁸ Autonomous Reanimation & Evacuation (AREVA) Research Institute and Innovation Center, The Geneva Foundation, San Antonio, TX, USA.
¹⁹ Defense Sciences Office, Defense Advanced Research Projects Agency, Arlington, VA, USA.
²⁰ Center for Translational Injury Research, Department of Surgery, McGovern Medical School at the University of Texas Health Science Center, Houston, TX, USA; CAG Center for Endotheliomics, Copenhagen University Hospital - Rigshospitalet, Copenhagen, Denmark; Department of Clinical Medicine, University of Copenhagen, Copenhagen, Denmark.
²¹ Center for Injury Science, University of Alabama at Birmingham, Birmingham, AL, USA; Division of Acute Care Surgery, Department of Surgery, University of Alabama at Birmingham, Birmingham, AL, USA.
²² RCSB Protein Data Bank, Rutgers, The State University of New Jersey, Piscataway, NJ, USA; RCSB Protein Data Bank, San Diego Supercomputer Center, University of California, San Diego, La Jolla, CA, USA; Institute for Quantitative Biomedicine, Rutgers, The State University of New Jersey, Piscataway, NJ, USA; Rutgers Cancer Institute, New Brunswick, NJ, USA.
²³ Center for Theoretical Biological Physics, Rice University, Houston, TX, USA; Department of Biosciences, Rice University, Houston, TX, USA; Department of Chemistry, Rice University, Houston, TX, USA; Department of Physics & Astronomy, Rice University, Houston, TX, USA.
²⁴ Rutgers Cancer Institute, Newark, NJ, USA; Division of Hematology/Oncology, Department of Medicine, Rutgers New Jersey Medical School, Newark, NJ, USA. Electronic address: wadih.arap@rutgers.edu.

PMID: 40609540
DOI: 10.1016/j.medj.2025.100638

Abstract

Background: Trauma is a leading cause of mortality, but injury-specific molecular targets remain largely unknown. We hypothesized that distinctive yet unrecognized tissue targets accessible to circulating ligands might emerge during trauma, thereby underscoring a trauma-related proteome.

Methods: We screened a peptide library to discover targets in a porcine model of major trauma: compound femur fracture with hemorrhagic shock. Bioinformatics yielded conserved motifs, and candidate receptors were affinity purified. In silico and in vitro approaches served to investigate possible associations between candidate receptors and calcium, a major component of skeletal muscle and bone. In vivo homing and molecular imaging (PET/MRI and SPECT/CT) studies of the most promising ligand peptide candidate were performed in the porcine model and were also confirmed in a corresponding rat model of major trauma. Optical methodologies and molecular dynamics simulations served to explore the molecular attributes of the ligand-receptor binding.

Findings: Nearly all molecular targets of the selected ligand peptides were calcium-dependent proteins, which become accessible upon trauma. We validated specific binding of homing peptides to these receptors in injured tissues, including CLRGFPALVC:CASQ1, CSEIGVRAC:HSP27, and CRQRPASGC:CALR. Notably, we determined that ligand peptide CRQRPASGC targets an injury-specific calcium-facilitated conformation of calreticulin, enabling specific molecular imaging of trauma.

Conclusions: We conceptually propose the term "traumome" for the functional receptor repertoire that becomes readily amenable for ligand-directed targeting upon major trauma. These preclinical findings pave the way toward clinic-ready targeted theragnostic approaches in the setting of trauma.

Funding: Major funding was provided by the Defense Advanced Research Projects Agency (DARPA).

Keywords: Pre-clinical research; acute trauma; calcium; calreticulin; compound fracture; in vivo screening; peptide library; phage display; receptor; shock; trauma-related proteome.

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Conflict of interest statement

Declaration of interests R.P. and W.A. are listed as inventors on a provisional patent application related to this technology and would be entitled to standard royalties if commercialization occurs. R.P., J.G.G., and W.A. are founders and equity shareholders of PhageNova Bio. R.P. is the Chief Scientific Officer of and serves as a paid consultant for PhageNova Bio. R.P. and W.A. are founders and equity shareholders of and serve as paid consultants for MBrace Therapeutics. F.I.S. is currently a full-time employee of MBrace Therapeutics. R.P. and W.A. have sponsored research agreements in place with both PhageNova Bio and MBrace Therapeutics. C.E.W. serves as a consultant for CellPhire Therapeutics, is a shareholder of Decisio Health, and receives funding from Grifols and Athersys. J.B.H. serves on the board of directors for Decisio Health, CCJ Medical Devices, QinFlow, Hemostatics, and Zibrio; he is a consultant for the Wake Forest Institute for Regenerative Medicine and Aspen Medical; he receives funding from CSL Behring; he is a co-inventor of the Junctional Emergency Treatment Tool; and he receives royalties from University of Texas (UT) Health. These arrangements are managed in accordance with the established institutional conflict-of-interest policies of the respective institutions. These conflicts of interest fall outside of the scope of this study.

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P30 CA016672/CA/NCI NIH HHS/United States

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Conformational ligand-directed targeting of calcium-dependent receptors in acute trauma

Affiliations

Conformational ligand-directed targeting of calcium-dependent receptors in acute trauma

Authors

Affiliations

Abstract

Conflict of interest statement

MeSH terms

Substances

Grants and funding