Classification and biomarker identification of prostate tissue from TRAMP mice with hyperpolarized 13C-SIRA

Anne B. Frahm; Deborah Hill; Sotirios Katsikis; Trygve Andreassen; Jan Henrik Ardenkjær-Larsen; Tone Frost Bathen; Siver Andreas Moestue; Pernille Rose Jensen; Mathilde Hauge Lerche

doi:10.1016/j.talanta.2021.122812

Classification and biomarker identification of prostate tissue from TRAMP mice with hyperpolarized ¹³C-SIRA

Anne B. Frahm
, Deborah Hill
, Sotirios Katsikis
, Trygve Andreassen
, Jan Henrik Ardenkjær-Larsen
, Tone Frost Bathen
, Siver Andreas Moestue
, Pernille Rose Jensen
, Mathilde Hauge Lerche^*

^*Corresponding author for this work

Norwegian University of Science and Technology

Research output: Contribution to journal › Journal article › Research › peer-review

122 Downloads (Orbit)

Abstract

Hyperpolarized ¹³C isotope resolved spectroscopy boosts NMR signal intensity, which improves signal detection and allows metabolic fluxes to be analyzed. Such hyperpolarized flux data may offer new approaches to tissue classification and biomarker identification that could be translated in vivo. Here we used hyperpolarized stable isotope resolved analysis (SIRA) to measure metabolite specific ¹³C isotopic enrichments in the central carbon metabolism of mouse prostate. Prostate and tumor tissue samples were acquired from transgenic adenocarcinomas of the mouse prostate (TRAMP) mice. Before euthanasia, mice were injected with [U–¹³C]glucose intraperitoneally (i.p.). Polar metabolite extracts were prepared, and hyperpolarized 1D-¹³C NMR spectra were obtained from normal prostate (n = 19) and cancer tissue (n = 19) samples. Binary classification and feature analysis was performed to make a separation model and to investigate differences between samples originating from normal and cancerous prostate tissue, respectively. Hyperpolarized experiments were carried out according to a standardized protocol, which showed a high repeatability (CV = 15%) and an average linewidth in the 1D-¹³C NMR spectra of 2 ± 0.5 Hz. The resolution of the hyperpolarized 1D-¹³C spectra was high with little signal overlap in the carbonyl region and metabolite identification was easily accomplished. A discrimination with 95% success rate could be made between samples originating from TRAMP mice prostate and tumor tissue based on isotopomers from uniquely identified metabolites. Hyperpolarized ¹³C-SIRA allowed detailed metabolic information to be obtained from tissue specimens. The positional information of ¹³C isotopic enrichments lead to easily interpreted features responsible for high predictive classification of tissue types. This analytical approach has matured, and the robust experimental protocols currently available allow systematic tracking of metabolite flux ex vivo.

Original language	English
Article number	122812
Journal	Talanta
Volume	235
Number of pages	23
ISSN	0039-9140
DOIs	https://doi.org/10.1016/j.talanta.2021.122812
Publication status	Published - 1 Dec 2021

Bibliographical note

Funding Information:
The authors gratefully acknowledge funding by the Danish National Research Foundation (Grant DNRF124 ).

Funding Information:
TRAMP mice breeding, housing, and genotyping was provided by the Comparative medicine Core Facility (CoMed), Norwegian University of Science and Technology (NTNU). CoMed is funded by the Faculty of Medicine at NTNU and Central Norway Regional Health Authority .

Funding Information:
Anne B. Frahm was supported by a travel and study grant from the Danish Cancer Society for a visiting stay at NTNU (Grant R235-A14222-19-S44 ).

Funding Information:
The tissue extraction procedures and 1 H -NMR experiments were performed at the MR Core Facility, Norwegian University of Science and Technology (NTNU). MR core facility is funded by the Faculty of Medicine and Health Sciences at NTNU and Central Norway Regional Health Authority .

Funding Information:
Hyperpolarized 1D- 13 C NMR data in this study was acquired with equipment partially funded by the Novo Nordisk Foundation ( NNF19OC0055825 ).

Funding Information:
The authors gratefully acknowledge funding by the Danish National Research Foundation (Grant DNRF124). Hyperpolarized 1D-13C NMR data in this study was acquired with equipment partially funded by the Novo Nordisk Foundation (NNF19OC0055825). The tissue extraction procedures and 1H -NMR experiments were performed at the MR Core Facility, Norwegian University of Science and Technology (NTNU). MR core facility is funded by the Faculty of Medicine and Health Sciences at NTNU and Central Norway Regional Health Authority. TRAMP mice breeding, housing, and genotyping was provided by the Comparative medicine Core Facility (CoMed), Norwegian University of Science and Technology (NTNU). CoMed is funded by the Faculty of Medicine at NTNU and Central Norway Regional Health Authority. The NMR center at DTU is acknowledged for the high-resolution 1H?13C HSQC and HMBC Spectrum acquired on their 800 MHz Avance II NMR spectrometer. Anne B. Frahm was supported by a travel and study grant from the Danish Cancer Society for a visiting stay at NTNU (Grant R235-A14222-19-S44).

Publisher Copyright:
© 2021

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Keywords

Dissolution dynamic nuclear polarization
Isotopic fingerprinting
Prostate cancer
Random forest
Stable isotope resolved analysis
Support vector machines

Access to Document

10.1016/j.talanta.2021.122812

FulltextAccepted author manuscript, 619 KB

OpenUrl availability

Check availability in DTU Findit

Cite this

@article{22273302ad484d90ac8430c2b83198ae,

title = "Classification and biomarker identification of prostate tissue from TRAMP mice with hyperpolarized 13C-SIRA",

abstract = "Hyperpolarized 13C isotope resolved spectroscopy boosts NMR signal intensity, which improves signal detection and allows metabolic fluxes to be analyzed. Such hyperpolarized flux data may offer new approaches to tissue classification and biomarker identification that could be translated in vivo. Here we used hyperpolarized stable isotope resolved analysis (SIRA) to measure metabolite specific 13C isotopic enrichments in the central carbon metabolism of mouse prostate. Prostate and tumor tissue samples were acquired from transgenic adenocarcinomas of the mouse prostate (TRAMP) mice. Before euthanasia, mice were injected with [U{\textendash}13C]glucose intraperitoneally (i.p.). Polar metabolite extracts were prepared, and hyperpolarized 1D-13C NMR spectra were obtained from normal prostate (n = 19) and cancer tissue (n = 19) samples. Binary classification and feature analysis was performed to make a separation model and to investigate differences between samples originating from normal and cancerous prostate tissue, respectively. Hyperpolarized experiments were carried out according to a standardized protocol, which showed a high repeatability (CV = 15\%) and an average linewidth in the 1D-13C NMR spectra of 2 ± 0.5 Hz. The resolution of the hyperpolarized 1D-13C spectra was high with little signal overlap in the carbonyl region and metabolite identification was easily accomplished. A discrimination with 95\% success rate could be made between samples originating from TRAMP mice prostate and tumor tissue based on isotopomers from uniquely identified metabolites. Hyperpolarized 13C-SIRA allowed detailed metabolic information to be obtained from tissue specimens. The positional information of 13C isotopic enrichments lead to easily interpreted features responsible for high predictive classification of tissue types. This analytical approach has matured, and the robust experimental protocols currently available allow systematic tracking of metabolite flux ex vivo.",

keywords = "Dissolution dynamic nuclear polarization, Isotopic fingerprinting, Prostate cancer, Random forest, Stable isotope resolved analysis, Support vector machines",

author = "Frahm, \{Anne B.\} and Deborah Hill and Sotirios Katsikis and Trygve Andreassen and Ardenkj{\ae}r-Larsen, \{Jan Henrik\} and Bathen, \{Tone Frost\} and Moestue, \{Siver Andreas\} and Jensen, \{Pernille Rose\} and Lerche, \{Mathilde Hauge\}",

note = "Funding Information: The authors gratefully acknowledge funding by the Danish National Research Foundation (Grant DNRF124 ). Funding Information: TRAMP mice breeding, housing, and genotyping was provided by the Comparative medicine Core Facility (CoMed), Norwegian University of Science and Technology (NTNU). CoMed is funded by the Faculty of Medicine at NTNU and Central Norway Regional Health Authority . Funding Information: Anne B. Frahm was supported by a travel and study grant from the Danish Cancer Society for a visiting stay at NTNU (Grant R235-A14222-19-S44 ). Funding Information: The tissue extraction procedures and 1 H -NMR experiments were performed at the MR Core Facility, Norwegian University of Science and Technology (NTNU). MR core facility is funded by the Faculty of Medicine and Health Sciences at NTNU and Central Norway Regional Health Authority . Funding Information: Hyperpolarized 1D- 13 C NMR data in this study was acquired with equipment partially funded by the Novo Nordisk Foundation ( NNF19OC0055825 ). Funding Information: The authors gratefully acknowledge funding by the Danish National Research Foundation (Grant DNRF124). Hyperpolarized 1D-13C NMR data in this study was acquired with equipment partially funded by the Novo Nordisk Foundation (NNF19OC0055825). The tissue extraction procedures and 1H -NMR experiments were performed at the MR Core Facility, Norwegian University of Science and Technology (NTNU). MR core facility is funded by the Faculty of Medicine and Health Sciences at NTNU and Central Norway Regional Health Authority. TRAMP mice breeding, housing, and genotyping was provided by the Comparative medicine Core Facility (CoMed), Norwegian University of Science and Technology (NTNU). CoMed is funded by the Faculty of Medicine at NTNU and Central Norway Regional Health Authority. The NMR center at DTU is acknowledged for the high-resolution 1H?13C HSQC and HMBC Spectrum acquired on their 800 MHz Avance II NMR spectrometer. Anne B. Frahm was supported by a travel and study grant from the Danish Cancer Society for a visiting stay at NTNU (Grant R235-A14222-19-S44). Publisher Copyright: {\textcopyright} 2021",

year = "2021",

month = dec,

day = "1",

doi = "10.1016/j.talanta.2021.122812",

language = "English",

volume = "235",

journal = "Talanta",

issn = "0039-9140",

publisher = "Elsevier",

}

TY - JOUR

T1 - Classification and biomarker identification of prostate tissue from TRAMP mice with hyperpolarized 13C-SIRA

AU - Frahm, Anne B.

AU - Hill, Deborah

AU - Katsikis, Sotirios

AU - Andreassen, Trygve

AU - Ardenkjær-Larsen, Jan Henrik

AU - Bathen, Tone Frost

AU - Moestue, Siver Andreas

AU - Jensen, Pernille Rose

AU - Lerche, Mathilde Hauge

N1 - Funding Information: The authors gratefully acknowledge funding by the Danish National Research Foundation (Grant DNRF124 ). Funding Information: TRAMP mice breeding, housing, and genotyping was provided by the Comparative medicine Core Facility (CoMed), Norwegian University of Science and Technology (NTNU). CoMed is funded by the Faculty of Medicine at NTNU and Central Norway Regional Health Authority . Funding Information: Anne B. Frahm was supported by a travel and study grant from the Danish Cancer Society for a visiting stay at NTNU (Grant R235-A14222-19-S44 ). Funding Information: The tissue extraction procedures and 1 H -NMR experiments were performed at the MR Core Facility, Norwegian University of Science and Technology (NTNU). MR core facility is funded by the Faculty of Medicine and Health Sciences at NTNU and Central Norway Regional Health Authority . Funding Information: Hyperpolarized 1D- 13 C NMR data in this study was acquired with equipment partially funded by the Novo Nordisk Foundation ( NNF19OC0055825 ). Funding Information: The authors gratefully acknowledge funding by the Danish National Research Foundation (Grant DNRF124). Hyperpolarized 1D-13C NMR data in this study was acquired with equipment partially funded by the Novo Nordisk Foundation (NNF19OC0055825). The tissue extraction procedures and 1H -NMR experiments were performed at the MR Core Facility, Norwegian University of Science and Technology (NTNU). MR core facility is funded by the Faculty of Medicine and Health Sciences at NTNU and Central Norway Regional Health Authority. TRAMP mice breeding, housing, and genotyping was provided by the Comparative medicine Core Facility (CoMed), Norwegian University of Science and Technology (NTNU). CoMed is funded by the Faculty of Medicine at NTNU and Central Norway Regional Health Authority. The NMR center at DTU is acknowledged for the high-resolution 1H?13C HSQC and HMBC Spectrum acquired on their 800 MHz Avance II NMR spectrometer. Anne B. Frahm was supported by a travel and study grant from the Danish Cancer Society for a visiting stay at NTNU (Grant R235-A14222-19-S44). Publisher Copyright: © 2021

PY - 2021/12/1

Y1 - 2021/12/1

N2 - Hyperpolarized 13C isotope resolved spectroscopy boosts NMR signal intensity, which improves signal detection and allows metabolic fluxes to be analyzed. Such hyperpolarized flux data may offer new approaches to tissue classification and biomarker identification that could be translated in vivo. Here we used hyperpolarized stable isotope resolved analysis (SIRA) to measure metabolite specific 13C isotopic enrichments in the central carbon metabolism of mouse prostate. Prostate and tumor tissue samples were acquired from transgenic adenocarcinomas of the mouse prostate (TRAMP) mice. Before euthanasia, mice were injected with [U–13C]glucose intraperitoneally (i.p.). Polar metabolite extracts were prepared, and hyperpolarized 1D-13C NMR spectra were obtained from normal prostate (n = 19) and cancer tissue (n = 19) samples. Binary classification and feature analysis was performed to make a separation model and to investigate differences between samples originating from normal and cancerous prostate tissue, respectively. Hyperpolarized experiments were carried out according to a standardized protocol, which showed a high repeatability (CV = 15%) and an average linewidth in the 1D-13C NMR spectra of 2 ± 0.5 Hz. The resolution of the hyperpolarized 1D-13C spectra was high with little signal overlap in the carbonyl region and metabolite identification was easily accomplished. A discrimination with 95% success rate could be made between samples originating from TRAMP mice prostate and tumor tissue based on isotopomers from uniquely identified metabolites. Hyperpolarized 13C-SIRA allowed detailed metabolic information to be obtained from tissue specimens. The positional information of 13C isotopic enrichments lead to easily interpreted features responsible for high predictive classification of tissue types. This analytical approach has matured, and the robust experimental protocols currently available allow systematic tracking of metabolite flux ex vivo.

AB - Hyperpolarized 13C isotope resolved spectroscopy boosts NMR signal intensity, which improves signal detection and allows metabolic fluxes to be analyzed. Such hyperpolarized flux data may offer new approaches to tissue classification and biomarker identification that could be translated in vivo. Here we used hyperpolarized stable isotope resolved analysis (SIRA) to measure metabolite specific 13C isotopic enrichments in the central carbon metabolism of mouse prostate. Prostate and tumor tissue samples were acquired from transgenic adenocarcinomas of the mouse prostate (TRAMP) mice. Before euthanasia, mice were injected with [U–13C]glucose intraperitoneally (i.p.). Polar metabolite extracts were prepared, and hyperpolarized 1D-13C NMR spectra were obtained from normal prostate (n = 19) and cancer tissue (n = 19) samples. Binary classification and feature analysis was performed to make a separation model and to investigate differences between samples originating from normal and cancerous prostate tissue, respectively. Hyperpolarized experiments were carried out according to a standardized protocol, which showed a high repeatability (CV = 15%) and an average linewidth in the 1D-13C NMR spectra of 2 ± 0.5 Hz. The resolution of the hyperpolarized 1D-13C spectra was high with little signal overlap in the carbonyl region and metabolite identification was easily accomplished. A discrimination with 95% success rate could be made between samples originating from TRAMP mice prostate and tumor tissue based on isotopomers from uniquely identified metabolites. Hyperpolarized 13C-SIRA allowed detailed metabolic information to be obtained from tissue specimens. The positional information of 13C isotopic enrichments lead to easily interpreted features responsible for high predictive classification of tissue types. This analytical approach has matured, and the robust experimental protocols currently available allow systematic tracking of metabolite flux ex vivo.

KW - Dissolution dynamic nuclear polarization

KW - Isotopic fingerprinting

KW - Prostate cancer

KW - Random forest

KW - Stable isotope resolved analysis

KW - Support vector machines

U2 - 10.1016/j.talanta.2021.122812

DO - 10.1016/j.talanta.2021.122812

M3 - Journal article

C2 - 34517669

AN - SCOPUS:85113278526

SN - 0039-9140

VL - 235

JO - Talanta

JF - Talanta

M1 - 122812

ER -