{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,20]],"date-time":"2025-10-20T18:54:54Z","timestamp":1760986494126,"version":"build-2065373602"},"reference-count":47,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2025,3,17]],"date-time":"2025-03-17T00:00:00Z","timestamp":1742169600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Dianne and Irving Kipnes Foundation"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Target modules (TMs), intermediate molecules required for UniCAR T-cell therapy, are promising molecules for immunotheranostic approaches. In the current work, we developed TMs containing a monomeric or dimeric form of the antagonist bombesin peptide (BBN2) and assessed their potential for diagnostic imaging using positron emission tomography (PET) as well as immunotherapy in combination with UniCAR T-cells to target and image GRPR expression in prostate cancer. Synthesized monomeric and dimeric BBN2 TMs retained binding to GRPR in vitro. Both BBN2 TMs specifically activated and redirected UniCAR T-cells to eradicate PC3 and LNCaP cancer cells with high efficiency and in a comparable manner. UniCAR T-cells retained a non-exhausted memory phenotype favorable to their persistence and fitness. The 68Ga-labeled BBN2 TMs showed proof-of-target towards GRPR in PC3 and LNCaP xenografts with similar uptake profiles for both BBN2 TMs in dynamic PET experiments. Clearance occurred exclusively through renal elimination. A tremendously increased in vivo metabolic stability of the BBN2 TMs was observed compared to their counterparts without E5B9. Both monomeric and dimeric BBN2 TMs represent novel and promising immunotheranostic tools for application in prostate cancer with exceptionally high in vivo metabolic stability.<\/jats:p>","DOI":"10.3390\/ijms26062686","type":"journal-article","created":{"date-parts":[[2025,3,17]],"date-time":"2025-03-17T06:36:23Z","timestamp":1742193383000},"page":"2686","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Tackling Prostate Cancer with Theranostic E5B9-Bombesin Target Modules (TMs): From Imaging to Treatment with UniCAR T-Cells"],"prefix":"10.3390","volume":"26","author":[{"given":"Liliana R.","family":"Loureiro","sequence":"first","affiliation":[{"name":"Institute of Radiopharmaceutical Cancer Research, Helmholtz-Zentrum Dresden-Rossendorf (HZDR), 01328 Dresden, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Susan","family":"Pike","sequence":"additional","affiliation":[{"name":"Department of Oncology, University of Alberta, Cross Cancer Institute, Edmonton, AB T6G 1Z2, Canada"},{"name":"Cancer Research Institute of Northern Alberta (CRINA), University of Alberta, Edmonton, AB T6G 2E1, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Melinda","family":"Wuest","sequence":"additional","affiliation":[{"name":"Department of Oncology, University of Alberta, Cross Cancer Institute, Edmonton, AB T6G 1Z2, Canada"},{"name":"Cancer Research Institute of Northern Alberta (CRINA), University of Alberta, Edmonton, AB T6G 2E1, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0000-8492-1525","authenticated-orcid":false,"given":"Cody N.","family":"Bergman","sequence":"additional","affiliation":[{"name":"Department of Oncology, University of Alberta, Cross Cancer Institute, Edmonton, AB T6G 1Z2, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kira R.","family":"J\u00d8rgensen","sequence":"additional","affiliation":[{"name":"Department of Oncology, University of Alberta, Cross Cancer Institute, Edmonton, AB T6G 1Z2, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ralf","family":"Bergmann","sequence":"additional","affiliation":[{"name":"Department of Biophysics and Radiation Biology, Semmelweis University, 1094 Budapest, Hungary"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anja","family":"Feldmann","sequence":"additional","affiliation":[{"name":"Institute of Radiopharmaceutical Cancer Research, Helmholtz-Zentrum Dresden-Rossendorf (HZDR), 01328 Dresden, Germany"},{"name":"German Cancer Consortium (DKTK), Partner Site Dresden, and German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany"},{"name":"National Center for Tumor Diseases Dresden (NCT\/UCC),  German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany"},{"name":"Faculty of Medicine and University Hospital Carl Gustav Carus, TU Dresden, 01307 Dresden, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Frank","family":"Wuest","sequence":"additional","affiliation":[{"name":"Department of Oncology, University of Alberta, Cross Cancer Institute, Edmonton, AB T6G 1Z2, Canada"},{"name":"Cancer Research Institute of Northern Alberta (CRINA), University of Alberta, Edmonton, AB T6G 2E1, Canada"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8029-5755","authenticated-orcid":false,"given":"Michael","family":"Bachmann","sequence":"additional","affiliation":[{"name":"Institute of Radiopharmaceutical Cancer Research, Helmholtz-Zentrum Dresden-Rossendorf (HZDR), 01328 Dresden, Germany"},{"name":"German Cancer Consortium (DKTK), Partner Site Dresden, and German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany"},{"name":"National Center for Tumor Diseases Dresden (NCT\/UCC),  German Cancer Research Center (DKFZ), 69120 Heidelberg, Germany"},{"name":"Faculty of Medicine and University Hospital Carl Gustav Carus, TU Dresden, 01307 Dresden, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,3,17]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"449","DOI":"10.1186\/s12967-023-04292-3","article-title":"Harnessing the potential of CAR-T cell therapy: Progress, challenges, and future directions in hematological and solid tumor treatments","volume":"21","author":"Dagar","year":"2023","journal-title":"J. Transl. Med."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"3494","DOI":"10.1182\/blood.V124.21.3494.3494","article-title":"Flexible Antigen-Specific Redirection of Human Regulatory T Cells Via a Novel Universal Chimeric Antigen Receptor System","volume":"124","author":"Koristka","year":"2014","journal-title":"Blood"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"e458","DOI":"10.1038\/bcj.2016.61","article-title":"Switching CAR T cells on and off: A novel modular platform for retargeting of T cells to AML blasts","volume":"6","author":"Cartellieri","year":"2016","journal-title":"Blood Cancer J."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"533","DOI":"10.1002\/jlcr.3727","article-title":"Theranostic CAR T cell targeting: A brief review","volume":"62","author":"Arndt","year":"2019","journal-title":"J. Label. Comp. Radiopharm."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1038\/s41408-018-0113-4","article-title":"Development of a novel target module redirecting UniCAR T cells to Sialyl Tn-expressing tumor cells","volume":"8","author":"Loureiro","year":"2018","journal-title":"Blood Cancer J."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"108584","DOI":"10.18632\/oncotarget.21017","article-title":"Development of novel target modules for retargeting of UniCAR T cells to GD2 positive tumor cells","volume":"8","author":"Mitwasi","year":"2017","journal-title":"Oncotarget"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"31368","DOI":"10.18632\/oncotarget.15572","article-title":"Retargeting of T lymphocytes to PSCA- or PSMA positive prostate cancer cells using the novel modular chimeric antigen receptor platform technology \u201cUniCAR\u201d","volume":"8","author":"Feldmann","year":"2017","journal-title":"Oncotarget"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/j.imlet.2019.05.003","article-title":"The UniCAR system: A modular CAR T cell approach to improve the safety of CAR T cells","volume":"211","author":"Bachmann","year":"2019","journal-title":"Immunol. Lett."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1016\/0014-4827(89)90038-4","article-title":"Identification of La ribonucleoproteins as a component of interchromatin granules","volume":"185","author":"Pfeifer","year":"1989","journal-title":"Exp. Cell Res."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Arndt, C., Fasslrinner, F., Loureiro, L.R., Koristka, S., Feldmann, A., and Bachmann, M. (2020). Adaptor car platforms\u2014Next generation of T cell-based cancer immunotherapy. Cancers, 12.","DOI":"10.3390\/cancers12051302"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"341","DOI":"10.1186\/s13046-023-02912-w","article-title":"Immunotheranostic target modules for imaging and navigation of UniCAR T-cells to strike FAP-expressing cells and the tumor microenvironment","volume":"42","author":"Loureiro","year":"2023","journal-title":"J. Exp. Clin. Cancer Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1186\/s13046-020-01572-4","article-title":"Extended half-life target module for sustainable UniCAR T-cell treatment of STn-expressing cancers","volume":"39","author":"Loureiro","year":"2020","journal-title":"J. Exp. Clin. Cancer Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"e1287246","DOI":"10.1080\/2162402X.2017.1287246","article-title":"A novel nanobody-based target module for retargeting of T lymphocytes to EGFR-expressing cancer cells via the modular UniCAR platform","volume":"6","author":"Albert","year":"2017","journal-title":"Oncoimmunology"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"25597","DOI":"10.18632\/oncotarget.25390","article-title":"From mono- to bivalent: Improving theranostic properties of target modules for redirection of UniCAR T cells against EGFR-expressing tumor cells in vitro and in vivo","volume":"9","author":"Albert","year":"2018","journal-title":"Oncotarget"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1659095","DOI":"10.1080\/2162402X.2019.1659095","article-title":"A theranostic PSMA ligand for PET imaging and retargeting of T cells expressing the universal chimeric antigen receptor UniCAR","volume":"8","author":"Arndt","year":"2019","journal-title":"Oncoimmunology"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"501","DOI":"10.1007\/s11307-017-1151-1","article-title":"Prostate Cancer Theranostics Targeting Gastrin-Releasing Peptide Receptors","volume":"20","author":"Baratto","year":"2018","journal-title":"Mol. Imaging Biol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1457","DOI":"10.1093\/annonc\/mdm058","article-title":"Gastrin-releasing peptide receptor as a molecular target in experimental anticancer therapy","volume":"18","author":"Cornelio","year":"2007","journal-title":"Ann. Oncol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"297","DOI":"10.1016\/j.cpet.2017.02.007","article-title":"Theranostic Prospects of Gastrin-Releasing Peptide Receptor-Radioantagonists in Oncology","volume":"12","author":"Maina","year":"2017","journal-title":"PET Clin."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1016\/j.cpet.2016.12.002","article-title":"From Bench to Bed: New Gastrin-Releasing Peptide Receptor-Directed Radioligands and Their Use in Prostate Cancer","volume":"12","author":"Maina","year":"2017","journal-title":"PET Clin."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1038\/nrurol.2013.42","article-title":"Targeting GRPR in urological cancers--from basic research to clinical application","volume":"10","author":"Mansi","year":"2013","journal-title":"Nat. Rev. Urol."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Li, H., Liu, Z., Yuan, L., Fan, K., Zhang, Y., Cai, W., and Lan, X. (2021). Radionuclide-Based Imaging of Breast Cancer: State of the Art. Cancers, 13.","DOI":"10.3390\/cancers13215459"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"921","DOI":"10.1097\/00001813-200411000-00001","article-title":"Targeting gastrin-releasing peptide receptors for cancer treatment","volume":"15","author":"Zhou","year":"2004","journal-title":"Anticancer Drugs"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Mansi, R., Nock, B.A., Dalm, S.U., Busstra, M.B., van Weerden, W.M., and Maina, T. (2021). Radiolabeled Bombesin Analogs. Cancers, 13.","DOI":"10.3390\/cancers13225766"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"557","DOI":"10.2967\/jnumed.115.168393","article-title":"Pilot Comparison of 68Ga-RM2 PET and 68Ga-PSMA-11 PET in Patients with Biochemically Recurrent Prostate Cancer","volume":"57","author":"Minamimoto","year":"2015","journal-title":"J. Nucl. Med."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"744","DOI":"10.2967\/jnumed.122.264448","article-title":"A Pilot Study of 68Ga-PSMA11 and 68Ga-RM2 PET\/MRI for Biopsy Guidance in Patients with Suspected Prostate Cancer","volume":"64","author":"Duan","year":"2023","journal-title":"J. Nucl. Med."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"75","DOI":"10.2967\/jnumed.116.178889","article-title":"Theranostic Perspectives in Prostate Cancer with the Gastrin-Releasing Peptide Receptor Antagonist NeoBOMB1: Preclinical and First Clinical Results","volume":"58","author":"Nock","year":"2017","journal-title":"J. Nucl. Med."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Gruber, L., Decristoforo, C., Uprimny, C., Hohenberger, P., Schoenberg, S.O., Orlandi, F., Mariani, M.F., Manzl, C., Kasseroler, M.T., and Tilg, H. (2022). Imaging Properties and Tumor Targeting of 68Ga-NeoBOMB1, a Gastrin-Releasing Peptide Receptor Antagonist, in GIST Patients. Biomedicines, 10.","DOI":"10.3390\/biomedicines10112899"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Wong, K., Sheehan-Dare, G., Nguyen, A., Ho, B., Liu, V., Lee, J., Brown, L., Dear, R., Chan, L., and Sharma, S. (2022). 64Cu-SAR-Bombesin PET-CT Imaging in the Staging of Estrogen\/Progesterone Receptor Positive, HER2 Negative Metastatic Breast Cancer Patients: Safety, Dosimetry and Feasibility in a Phase I Trial. Pharmaceuticals, 15.","DOI":"10.3390\/ph15070772"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1025","DOI":"10.1016\/j.nucmedbio.2013.07.005","article-title":"Synthesis and radiopharmacological evaluation of a high-affinity and metabolically stabilized 18F-labeled bombesin analogue for molecular imaging of gastrin-releasing peptide receptor-expressing prostate cancer","volume":"40","author":"Richter","year":"2013","journal-title":"Nucl. Med. Biol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1347","DOI":"10.1021\/acs.molpharmaceut.5b00970","article-title":"Metabolically Stabilized (68)Ga-NOTA-Bombesin for PET Imaging of Prostate Cancer and Influence of Protease Inhibitor Phosphoramidon","volume":"13","author":"Richter","year":"2016","journal-title":"Mol. Pharm."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"74","DOI":"10.1016\/j.nucmedbio.2020.10.005","article-title":"A comparative PET imaging study of 44gSc- and 68Ga-labeled bombesin antagonist BBN2 derivatives in breast and prostate cancer models","volume":"90","author":"Ferguson","year":"2020","journal-title":"Nucl. Med. Biol."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Benichou, G., Gonzalez, B., Marino, J., Ayasoufi, K., and Valujskikh, A. (2017). Role of Memory T Cells in Allograft Rejection and Tolerance. Front. Immunol., 8.","DOI":"10.3389\/fimmu.2017.00170"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"e150807","DOI":"10.1172\/JCI150807","article-title":"CAR T cell manufacturing from naive\/stem memory T lymphocytes enhances antitumor responses while curtailing cytokine release syndrome","volume":"132","author":"Arcangeli","year":"2022","journal-title":"J. Clin. Investig."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"L\u00f3pez-Cantillo, G., Urue\u00f1a, C., Camacho, B.A., and Ram\u00edrez-Segura, C. (2022). CAR-T Cell Performance: How to Improve Their Persistence?. Front. Immunol., 13.","DOI":"10.3389\/fimmu.2022.878209"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"270","DOI":"10.1016\/j.trecan.2022.12.008","article-title":"T cell exhaustion in malignant gliomas","volume":"9","author":"Watowich","year":"2023","journal-title":"Trends Cancer"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"3938","DOI":"10.1007\/s00259-022-05831-8","article-title":"Diverse PSMA expression in primary prostate cancer: Reason for negative [68Ga]Ga-PSMA PET\/CT scans? Immunohistochemical validation in 40 surgical specimens","volume":"49","author":"Cytawa","year":"2022","journal-title":"Eur. J. Nucl. Med. Mol. Imaging"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"75","DOI":"10.4155\/fdd-2022-0005","article-title":"Overcoming the shortcomings of peptide-based therapeutics","volume":"4","author":"Lamers","year":"2022","journal-title":"Future Drug Discov."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1175","DOI":"10.1016\/j.drudis.2018.02.012","article-title":"Approved CAR T cell therapies: Ice bucket challenges on glaring safety risks and long-term impacts","volume":"23","author":"Zheng","year":"2018","journal-title":"Drug Discov. Today"},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Rangger, C., and Haubner, R. (2020). Radiolabelled Peptides for Positron Emission Tomography and Endoradiotherapy in Oncology. Pharmaceuticals, 13.","DOI":"10.3390\/ph13020022"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1053\/j.semnuclmed.2017.05.007","article-title":"Clinical Applications of Radiolabeled Peptides for PET","volume":"47","author":"Jackson","year":"2017","journal-title":"Semin. Nucl. Med."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Prasad, V. (2024). Theranostic Radiopeptides in Nuclear Oncology: Design, Preclinical Screening, and Clinical Translation. Beyond Becquerel and Biology to Precision Radiomolecular Oncology: Festschrift in Honor of Richard P Baum, Springer. [1st ed.].","DOI":"10.1007\/978-3-031-33533-4"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1081","DOI":"10.1007\/s00726-010-0546-y","article-title":"Peptide heterodimers for molecular imaging","volume":"41","author":"Yan","year":"2011","journal-title":"Amino Acids"},{"key":"ref_43","doi-asserted-by":"crossref","unstructured":"Hedhli, J., Czerwinski, A., Schuelke, M., P\u0142oska, A., Sowinski, P., Hood, L., Mamer, S.B., Cole, J.A., Czaplewska, P., and Banach, M. (2017). Synthesis, Chemical Characterization and Multiscale Biological Evaluation of a Dimeric-cRGD Peptide for Targeted Imaging of \u03b1 V \u03b2 3 Integrin Activity. Sci. Rep., 7.","DOI":"10.1038\/s41598-017-03224-8"},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Judmann, B., Braun, D., W\u00e4ngler, B., Schirrmacher, R., Fricker, G., and W\u00e4ngler, C. (2020). Current State of Radiolabeled Heterobivalent Peptidic Ligands in Tumor Imaging and Therapy. Pharmaceuticals, 13.","DOI":"10.3390\/ph13080173"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"121","DOI":"10.2967\/jnumed.113.129411","article-title":"\u201cTo Serve and Protect\u201d: Enzyme Inhibitors as Radiopeptide Escorts Promote Tumor Targeting","volume":"55","author":"Nock","year":"2014","journal-title":"J. Nucl. Med."},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Rethi-Nagy, Z., Abraham, E., Udvardy, K., Klement, E., Darula, Z., Pal, M., Katona, R.L., Tubak, V., Pali, T., and Kota, Z. (2022). STABILON, a Novel Sequence Motif That Enhances the Expression and Accumulation of Intracellular and Secreted Proteins. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23158168"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"214","DOI":"10.1034\/j.1399-3011.2003.00085.x","article-title":"Using protein-based motifs to stabilize peptides","volume":"62","author":"Walker","year":"2003","journal-title":"J. Pept. Res."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/6\/2686\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T16:54:53Z","timestamp":1760028893000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/6\/2686"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,3,17]]},"references-count":47,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2025,3]]}},"alternative-id":["ijms26062686"],"URL":"https:\/\/doi.org\/10.3390\/ijms26062686","relation":{},"ISSN":["1422-0067"],"issn-type":[{"type":"electronic","value":"1422-0067"}],"subject":[],"published":{"date-parts":[[2025,3,17]]}}}