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journal article·2026-06-23Open access

Hyperpolarization of [1‐ 13 C]Ketoisocaproate‐d 2 by Reversible Exchange with Parahydrogen Enables Profiling of Branched‐Chain‐Amino‐Acid Metabolism in Cellulo and in Vivo

Stefan Petersen, Philipp R. Groß, Paul M. Schmidt, Henri de Maissin, Asitan Rittinger, Robert Willing, Adriana Sacristán‐Martín, Lisa Heß, Jule Koch, Sebastian Lucas, Maxim Zaitsev, Dominik von Elverfeldt, Martin Grashei, Franz Schilling, Jan‐Bernd Hövener, André F. Martins, Max von Delius, Thomas Reinheckel, Andreas B. Schmidt

Advanced Science
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Abstract

ABSTRACT Hyperpolarized 1 3 C magnetic resonance imaging (MRI) is the only method to image metabolic fluxes in real time, non‐invasively, and in vivo. To date, however, most studies have used [1‐ 1 3 C]pyruvate and dynamic nuclear polarization (dDNP). Here, we establish efficient hyperpolarization (HP) of protio and partially‐deuterated [1‐ 1 3 C]ketoisocaproate (KIC) using Spin‐Lock‐Induced‐Crossing‐Signal Amplification by Reversible Exchange (SLIC‐SABRE), a high‐throughput, uncomplex and low‐cost method based on parahydrogen. We demonstrate 13 C polarization up to ≈28% and T 1 relaxation times > 200 s at 1 T in methanol‐d 4 . A rapid purification procedure allowed us to obtain biocompatible formulations with ≈11% 13 C polarization at the time of injection, sufficient for in cellulo and in vivo studies. We found that branched‐chain‐amino‐acid transaminase (BCAT) activity leads to HP [1‐ 1 3 C]leucine formation exclusively in BCAT1‐high MDA‐MB231 breast‐cancer cells, but not in BCAT1‐low MCF7 or PyB6 cells. In a proof‐of‐concept in vivo experiment, 13 C magnetic resonance spectroscopy imaging of the healthy mouse brain detected [1‐ 13 C]leucine formation after intravenous injection of SABRE [1‐ 13 C]KIC‐d 2 . Our results demonstrate that [1‐ 13 C]KIC‐d 2 provides a sensitive readout of BCAT1‐dependent metabolism and that SLIC‐SABRE can rapidly generate this probe for 13 C MRI, extending accessible parahydrogen‐based hyperpolarization to amino‐acid pathways relevant to cancer biology and chemoresistance.
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Research context

Biomaterials

OpenAlex topics: Advanced NMR Techniques and Applications · Advanced MRI Techniques and Applications · Electron Spin Resonance Studies

SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.

Affiliations

Division of Medical Physics Department of Radiology Medical Center ‐ University of Freiburg Faculty of Medicine ‐ University of Freiburg Freiburg Germany

German Cancer Consortium (DKTK) partner site Freiburg a Partnership Between DKFZ and University Medical Center Freiburg Heidelberg Germany

Faculty of Biology Albert Ludwigs University Freiburg Freiburg Germany

Institute of Organic Chemistry and Center for Integrated Quantum Science and Technology (IQST) University of Ulm Ulm Germany

Institute of Molecular Medicine and Cell Research Faculty of Medicine University of Freiburg Freiburg Germany

NVision Imaging Technologies GmbH Ulm Germany

Technical University of Munich TUM School of Medicine and Health Department of Nuclear Medicine TUM University Hospital Munich Germany

German Cancer Consortium (DKTK), partner site Munich a Partnership Between DKFZ and TUM Heidelberg Germany

Section Biomedical Imaging Molecular Imaging North Competence Center MOINCC Department of Radiology and Neuroradiology University Hospital Schleswig Holstein Kiel University Kiel Germany

Werner Siemens Imaging Center Department of Preclinical Imaging and Radiopharmacy Eberhard Karls University Tübingen Tübingen Germany

Cluster of Excellence iFIT (EXC 2180) “Image‐Guided and Functionally Instructed Tumor Therapies” Eberhard Karls University Tübingen Tübingen Germany

German Cancer Consortium (DKTK), partner site Tübingen a Partnership Between DKFZ and University Hospital Tübingen Heidelberg Germany

Centre for Biological Signalling Studies BIOSS University of Freiburg Freiburg Germany

Funding

Deutsche Forschungsgemeinschaft

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