Public Abstract

DE-SC0025288: Single Cell level elemental signatures of low dose radiation exposures in mammalian model systems

Award Status: Active

Institution: Northwestern University, Chicago, IL
UEI: KG76WYENL5K1
DUNS:

Most Recent Award Date: 09/11/2024
Number of Support Periods: 1
PM: Kulkarni, Resham

Current Budget Period: 09/01/2024 - 08/31/2025
Current Project Period: 09/01/2024 - 08/31/2027
PI: Woloschak, Gayle

Supplement Budget Period: N/A

Public Abstract

Title: Single Cell level elemental signatures of low dose radiation exposures in mammalian model systems Principal Investigator: Gayle Woloschak, Professor, Northwestern University (NU) Senior/Key Personnel: Tatjana Paunesku, Stuart Stock, Ching Man Wai, Priyam Patel, Zequn Sun, Demirkan Gürsel, Craig Horbinski, Qiaoling Jin and Lori Snyder, Northwestern University (NU) Ahtesham Ullah Khan, Northwestern Medical Hospital (NMH) Dorthe Schaue, University of California Los Angeles (UCLA) Stefan Vogt, Mathew Cherukara, Francesco De Carlo, Si Chen, Barry Lai and Olga Antipova, Advanced Photon Source, Argonne National Laboratory (APS/ANL) Rebecca Abergel, Lawrence Berkeley National Laboratory (LBNL) Project objectives This proposal will test the hypothesis that low dose radiation exposures lead to changes in vascularization of normal tissues, a health outcome that may not cause life shortening, while at the same time it may cause changes in disease incidence. The best way to study changes in vascularization is to work with low dose exposed model organisms that have been allowed to live out their entire lifespan. The PI’s laboratory maintains and curates an archive of data and tissues from beagle dogs exposed to low doses of radiation. Among the animals that received low LET beta emitters are beagles from the University of California Davis that ingested radioactive strontium-90. No significant life shortening was found in lowest dose groups –archival samples from these animals will be used for these studies. Project description and methods Because strontium accumulates in bones, especially bones of the skull, we have selected brain for study of microvasculature. Dr. Paunesku (NU) will identify and extract archival FFPE beagle brain tissues from NURA animals exposed to strontium-90 in doses that constitute chronic low dose radiation exposures. Dr. Abergel (LBNL) will inject 40 mice with low doses of Sr-90 and, after a month-long post-exposure, isolate brain tissues plus 25 control brains. Dr. Khan (NU) will perform Monte Carlo simulations to provide detailed dose calculations for animal brains. Dr. Stock (NU) will use LXCT instrument to evaluate the vasculature in beagle FFPE brain tissues. Dr. De Carlo (APS, 2BMB) will use the sXCT instrument developed by his team for 3D phase contrast X-ray imaging of rodent brains with 1 µm volume elements. Beagle brain vasculature 3D maps from synchrotron and laboratory CT will be examined by Dr. De Carlo and Dr. Stock with support from Dr. Iruela-Arispe (NU) who is an expert on vessel abnormalities. The focus will be on detecting “sprouting angiogenesis” to select subsections of FFPE samples of interest for further studies. Next steps of sample processing will include optical imaging and pathology evaluation that will be done by other members of the NU team, including Drs. Paunesku, Gürsel and Horbinski who have worked together on elemental imaging of human brain samples in the past (Kumthekar, Ko et al. 2021). Dr. Jin will assist in this effort as well. In addition, Dr. Schaue (UCLA) will assess these samples from a radiobiology-immunology perspective, focusing on presence and distribution of macrophages and their possible role in new angiogenesis. The samples and sample portions with the most promising pathology features will be “reformatted” to fit different x-ray spectroscopy instruments at the APS. These samples will be subjected to elementalomic evaluation conducted with support and in collaboration with the investigators from Sector 8 and 2 beamlines (Drs. Lai, Chen, Antipova and Jin, APS, 8BMB, 2IDE, 2IDD, BioNanoprobe). Elementalomic changes in brain samples will be evaluated with special attention on copper – an element that is critically involved in angiogenesis as shown in earlier 2D elemental studies done by the members of this team e.g. (Finney, Mandava et al. 2007, Qin, Toursarkissian et al. 2011, Heuberger, Harankhedkar et al. 2019). Micro RNAs in archival tissues will be studied using spatial transcriptomics resources at NU in collaboration with Dr. Wai. Xenium In Situ System from 10x Genomics will be used for evaluation of archival samples(NU), while new mouse samples will be run on both Visium and Xenium In Situ Systems. Dr. Patel (NU). Dr. Sun (NU) will be involved in the statistical data analyses for all different parts of the work on this proposal, including the miRNA transcriptomics data. Spatial transcriptomic data from archival and fresh tissues will be analyzed with the primary focus on miRNAs and gene expression pathways that correlate with angiogenesis, vascular endothelial cell activity, and copper utilization as well as macrophage activity. AI/ML work led by Dr. Cherukara (APS) will primarily support the extensive computational demands of data acquisition, 2D and 3D image reconstruction and image correlations. In addition, this team and the NU bioinformatics and statistics team will collaborate to analyze and correlate the complete data – including spatial miRNA expression, elementalomic information in 2D and 3D, micro-CT information and histopathology information. This work will for a roadmap for characterization of vascular sprouting anomalies in the context of radiation-induced brain complications found in archival canine samples. Potential impact of the project New computational tools will be developed to support high dimensional data acquisition and processing allowing us to link the x-ray generated 2D and 3D images with the data from visible light imaging and spatial transcriptomics at the level of single cells. This work will deepen our understanding of the effects of low dose radiation exposures, inspire development of new research pipelines and generate a wealth of data for future analyses.

Title: Single Cell level elemental signatures of low dose radiation exposures in mammalian model systems

Principal Investigator: Gayle Woloschak, Professor, Northwestern University (NU)

Senior/Key Personnel:

Tatjana Paunesku, Stuart Stock, Ching Man Wai, Priyam Patel, Zequn Sun, Demirkan Gürsel, Craig Horbinski, Qiaoling Jin and Lori Snyder, Northwestern University (NU)

Ahtesham Ullah Khan, Northwestern Medical Hospital (NMH)

Dorthe Schaue, University of California Los Angeles (UCLA)

Stefan Vogt, Mathew Cherukara, Francesco De Carlo, Si Chen, Barry Lai and Olga Antipova, Advanced Photon Source, Argonne National Laboratory (APS/ANL)

Rebecca Abergel, Lawrence Berkeley National Laboratory (LBNL)

Project objectives

This proposal will test the hypothesis that low dose radiation exposures lead to changes in vascularization of normal tissues, a health outcome that may not cause life shortening, while at the same time it may cause changes in disease incidence. The best way to study changes in vascularization is to work with low dose exposed model organisms that have been allowed to live out their entire lifespan. The PI’s laboratory maintains and curates an archive of data and tissues from beagle dogs exposed to low doses of radiation. Among the animals that received low LET beta emitters are beagles from the University of California Davis that ingested radioactive strontium-90. No significant life shortening was found in lowest dose groups –archival samples from these animals will be used for these studies.

Project description and methods

Because strontium accumulates in bones, especially bones of the skull, we have selected brain for study of microvasculature. Dr. Paunesku (NU) will identify and extract archival FFPE beagle brain tissues from NURA animals exposed to strontium-90 in doses that constitute chronic low dose radiation exposures. Dr. Abergel (LBNL) will inject 40 mice with low doses of Sr-90 and, after a month-long post-exposure, isolate brain tissues plus 25 control brains. Dr. Khan (NU) will perform Monte Carlo simulations to provide detailed dose calculations for animal brains. Dr. Stock (NU) will use LXCT instrument to evaluate the vasculature in beagle FFPE brain tissues. Dr. De Carlo (APS, 2BMB) will use the sXCT instrument developed by his team for 3D phase contrast X-ray imaging of rodent brains with 1 µm volume elements. Beagle brain vasculature 3D maps from synchrotron and laboratory CT will be examined by Dr. De Carlo and Dr. Stock with support from Dr. Iruela-Arispe (NU) who is an expert on vessel abnormalities. The focus will be on detecting “sprouting angiogenesis” to select subsections of FFPE samples of interest for further studies. Next steps of sample processing will include optical imaging and pathology evaluation that will be done by other members of the NU team, including Drs. Paunesku, Gürsel and Horbinski who have worked together on elemental imaging of human brain samples in the past (Kumthekar, Ko et al. 2021). Dr. Jin will assist in this effort as well. In addition, Dr. Schaue (UCLA) will assess these samples from a radiobiology-immunology perspective, focusing on presence and distribution of macrophages and their possible role in new angiogenesis. The samples and sample portions with the most promising pathology features will be “reformatted” to fit different x-ray spectroscopy instruments at the APS. These samples will be subjected to elementalomic evaluation conducted with support and in collaboration with the investigators from Sector 8 and 2 beamlines (Drs. Lai, Chen, Antipova and Jin, APS, 8BMB, 2IDE, 2IDD, BioNanoprobe). Elementalomic changes in brain samples will be evaluated with special attention on copper – an element that is critically involved in angiogenesis as shown in earlier 2D elemental studies done by the members of this team e.g. (Finney, Mandava et al. 2007, Qin, Toursarkissian et al. 2011, Heuberger, Harankhedkar et al. 2019). Micro RNAs in archival tissues will be studied using spatial transcriptomics resources at NU in collaboration with Dr. Wai. Xenium In Situ System from 10x Genomics will be used for evaluation of archival samples(NU), while new mouse samples will be run on both Visium and Xenium In Situ Systems. Dr. Patel (NU). Dr. Sun (NU) will be involved in the statistical data analyses for all different parts of the work on this proposal, including the miRNA transcriptomics data. Spatial transcriptomic data from archival and fresh tissues will be analyzed with the primary focus on miRNAs and gene expression pathways that correlate with angiogenesis, vascular endothelial cell activity, and copper utilization as well as macrophage activity. AI/ML work led by Dr. Cherukara (APS) will primarily support the extensive computational demands of data acquisition, 2D and 3D image reconstruction and image correlations. In addition, this team and the NU bioinformatics and statistics team will collaborate to analyze and correlate the complete data – including spatial miRNA expression, elementalomic information in 2D and 3D, micro-CT information and histopathology information. This work will for a roadmap for characterization of vascular sprouting anomalies in the context of radiation-induced brain complications found in archival canine samples.

Potential impact of the project

New computational tools will be developed to support high dimensional data acquisition and processing allowing us to link the x-ray generated 2D and 3D images with the data from visible light imaging and spatial transcriptomics at the level of single cells. This work will deepen our understanding of the effects of low dose radiation exposures, inspire development of new research pipelines and generate a wealth of data for future analyses.