SBIR-STTR Award

PhotoSound Technologies, Inc. proposes to develop a novel imaging modality for characterization and pre- clinical research of murine models. The technology will be capable of three-dimensional functional and molecular imaging of fluorescent labels and reporter genes mapped with high fidelity over robust anatomical structures, such as skin, central and peripheral vasculature, and internal organs. The developed instrument could be used in broad spectrum of pre-clinical research including cancer, toxicology, tissue engineering and regeneration, cardiovascular and developmental biology. Optical in vivo imaging methods (fluorescence and bioluminescence) found great popularity among researchers as affordable, convenient, and very sensitive molecular imaging tools for pre-clinical studies and development of animal models. However, their stand-alone application is impeded by poor spatial resolution and limitations imposed by two-dimensionality of the images. A high-resolution in vivo 3D imaging method, which could be easily integrated with optical imaging in a single instrument, would have a great impact on the entire field of small animal research. Photoacoustic tomography is an emerging biomedical imaging modality that has all those requirements: (1) 150-500 µm resolution of 3D whole body images; (2) Ability to use the same instrumentation for excitation of fluorescence and generation of photoacoustic effect; (3) 3D scans in less than 1 minute. However, its in vivo sensitivity to detection of fluorophores is inferior as compared to regular fluorescence techniques. Our proposal is based on pioneering co-registered integration of fluorescence and photoacoustic modalities in a single compact 3D configuration (PAFT-3D) defeating shortcomings of each individual technology. The Phase I project is organized in three specific aims assessing feasibility of: (1) Technological implementation for co-registered 3D photoacoustic tomography and fluorescence in a single compact instrument for imaging live anesthetized mice; (2) Imaging fluorophores and anatomical structures of mice with resolution exceeding that of established fluorescence imaging; (3) Providing imaging information valuable for development and characterization of pre-clinical animal models with case study focused on murine models of metastatic breast cancer. Phase II will be focused on development and in vivo validation of a commercial instrument with sensitivity and imaging procedures optimized for various areas of animal model development and preclinical research. Ultimate commercial system will enable in vivo visualization and analysis of native hemoglobin, fluorophores, nanoparticles, and other photosensitive constructs used for tracking, mapping, and longitudinal studies. The value proposition of the PAFT-3D will be built not only around uniqueness and technological superiority of the product, but also by making it affordable for a broad population of research groups and centers.

Public Health Relevance Statement:
Well-established, cheap, and convenient rodent models of human disease currently represent the golden standard and the first step of in vivo pre-clinical studies. Development and characterization of such models requires non-invasive functional and molecular imaging techniques, which currently have significant deficiencies in spatial resolution, two-dimensionality of the images, and lack of convenient and cheap instrumentation providing co-registered anatomical referencing. This project proposes development of a compact 3D imaging solution integrating molecular, functional, and anatomical imaging with unparallel combination of sensitivity and resolution, which could be employed in broad spectrum of pre-clinical research areas including cancer, toxicology, tissue engineering and regeneration, cardiovascular and developmental medicine and biology. 1

Project Terms:
Anatomy Qualifier; Anatomical Sciences; Anatomic structures; Anatomic Structures and Systems; Anatomic Structure, System, or Substance; Anatomic Sites; Anatomic; Anatomy; Animal Research; Animal Experimental Use; Animal Experimentation; Biology; Biomedical Research; vascular; Blood Vessels; neoplasm/cancer; malignancy; Malignant Tumor; Cancers; Malignant Neoplasms; circulatory system; Heart Vascular; Cardiovascular Organ System; Cardiovascular Body System; Cardiovascular; Cardio-vascular; Cardiovascular system; Developmental Biology; Engineering; Fluorescence; Gold; Hemoglobin; instrumentation; Kidney; renal; Kidney Urinary System; Light; Photoradiation; Lipids; Longitudinal Studies; long-term study; Maps; Medicine; Melanins; Methodology; Mus; Murine; Mice Mammals; Mice; Neoplasm Metastasis; tumor cell metastasis; cancer metastasis; Secondary Tumor; Secondary Neoplasm; Metastatic Tumor; Metastatic Neoplasm; Metastatic Mass; Metastatic Lesion; Metastasize; Metastasis; Research; research and development; R&D; R & D; Development and Research; Research Personnel; Researchers; Investigators; Spleen; Spleen Reticuloendothelial System; Technology; Testing; Toxicology; Trees; ultrasound scanning; ultrasound imaging; ultrasound; sound measurement; sonography; sonogram; diagnostic ultrasound; Ultrasound Test; Ultrasound Medical Imaging; Ultrasound Diagnosis; Ultrasonogram; Ultrasonic Imaging; Medical Ultrasound; Echotomography; Echography; Ultrasonography; Hydrogen Oxide; Water; Generations; Imaging Technics; Imaging Procedures; Imaging Techniques; case report; Case Study; Bioluminescence; Visualization; Imagery; base; Organ; Label; Procedures; whole body scanning; whole body imaging; Peripheral; Area; Phase; Biological; Reporter Genes; Individual; Measurement; cancer cell; Malignant Cell; instrument; Photosensitivity; sun sensitivity; Photosensitiveness due to sun; Dimensions; Scanning; Techniques; System; Inferior; Tissue Engineering; engineered tissue; chromophore; experience; fluorophore; Animal Model; model organism; model of animal; Animal Models and Related Studies; Structure; novel; Modality; Devices; Code; Coding System; Modeling; bioimaging; biomedical imaging; bio-imaging; Three-Dimensional Imaging; Three Dimensional Medical Imaging; 3D imaging; 3-D Imaging; Skin; Three-Dimensional Image; 3D images; 3D image; 3-D image; 3-D Images; Quantum Dots; Q-Dot; imaging modality; imaging method; image-based method; Address; Breast Cancer Model; mammary tumor model; mammary cancer model; Breast tumor model; Data; Detection; Imaging Device; Imaging Tool; Imaging Instrument; Population Research; Population-level research; Population-based research; Resolution; in vivo; physiological imaging; Physiologic Imaging; Functional Imaging; Optical Methods; Rodent Model; Small Business Innovation Research; SBIR; Small Business Innovation Research Grant; Validation; Molecular; Process; Sentinel Node; Sentinel Lymph Node; developmental; Development; molecule imaging; molecular imaging; regenerating damaged tissue; regenerate new tissue; tissue regeneration; imaging; Image; preclinical; pre-clinical; pre-clinical study; preclinical study; cost; optic imaging; optical imaging; reconstruction; designing; design; nano particle; nanoparticle; Outcome; Imaging technology; Carbon nanoparticle; innovation; innovative; innovate; data acquisition; pre-clinical research; preclinical research; two-dimensional; 2-dimensional; graphical user interface; graphic user interface; Graphical interface; human disease; mouse model; murine model; fluorescence imaging; fluorescent imaging; prototype; commercialization; animal model development; plasmonics; photoacoustic imaging; optoacoustic imaging; in vivo imaging; Metastatic breast cancer; imaging system; high resolution imaging; anatomic imaging; anatomical imaging; optoacoustic tomography; photoacoustic tomography; clinical development; in vivo optical imaging; imaging approach

PhotoSound Technologies, Inc. proposes to develop a novel imaging modality for characterization and preclinical research of small animal models. The technology will be capable of three-dimensional functional and molecular imaging of fluorescent labels and reporter genes mapped with high fidelity over robust anatomical structures, such as skin, central and peripheral vasculature, and internal organs. The Phase II commercial instrument is designed to perform high-throughput whole body imaging of rodent models. It could be used in broad spectrum of preclinical research applications including cancer, toxicology, tissue engineering and regeneration, cardiovascular and developmental biology. In addition to qualitatively superior performance characteristics, the proposed multimodality imaging platform will significantly reduce space and funds required to house and operate a whole-body imaging platform at an animal research facility. Optical in vivo imaging methods (fluorescence and bioluminescence) found great popularity among researchers as affordable, convenient, and very sensitive molecular imaging tools for preclinical studies and development of animal models. However, their stand-alone application is impeded by poor spatial resolution and limitations imposed by two-dimensionality of the images. A fast and high-resolution in vivo 3D imaging method, which could be easily integrated with optical imaging in a single instrument, would have a great impact on the entire field of small animal research. Photoacoustic tomography is an emerging whole body 3D imaging modality capable of 200-500 µm resolution. It can also use the same components for excitation of fluorescence and generation of photoacoustic effect. However, its in vivo application for detection of fluorophores is impeded by strong background generated by native blood. Also, there are no commercial or research photoacoustic whole- body imaging instrument that could work with high-throughput imaging procedures (<5 min per animal). Our proposal shows a way to defeat shortcomings of each individual technology and enable fast high-resolution whole body 3D imaging of fluorescent biomarkers by integrating robotic scanning and multi-view orthogonal fluorescence and photoacoustics in a single co-registered modality (PAFT). The Phase II project is focused on development of a commercial PAFT instrument and is organized in three specific aims: (1) Develop, fabricate, and assess the performance of a commercial PAFT instrument; (2) Develop and implement high-throughput PAFT image reconstruction method; (3) Field-test the commercial PAFT instrument. Ultimate commercial system will enable high-throughput in vivo 3D visualization and analysis of native hemoglobin, fluorophores, nanoparticles, and other photosensitive constructs used for molecular and functional mapping and longitudinal studies. Such an instrument would alleviate subjective interpretation of missing and misregistered imaging data and would streamline the preclinical phase of development, leading to a higher success rate and lower costs of drug development.

Public Health Relevance Statement:
Well-established, cheap, and convenient rodent models of human disease currently represent the golden standard and the first step of in vivo preclinical studies. Development, characterization, and use of such models requires high-throughput whole body functional and molecular imaging techniques, which currently have significant deficiencies in spatial resolution, two-dimensionality of the images, or lack of multimodality co- registration and anatomical referencing. This project proposes development of a high-throughput whole body 3D imaging solution integrating molecular, functional, and anatomical imaging with unparallel spatial resolution, which could be efficiently employed in spatially constrained environments of shared animal facilities and contract research organizations (CROs). 1

Project Terms:
Algorithms; Anatomy; Anatomic; Anatomic Sites; Anatomic structures; Anatomical Sciences; Animal Experimentation; Animal Experimental Use; Animal Research; Animals; Biomedical Research; Blood; Blood Reticuloendothelial System; Malignant Neoplasms; Cancers; Malignant Tumor; malignancy; neoplasm/cancer; Cardiovascular system; Cardiovascular; Cardiovascular Body System; Cardiovascular Organ System; Heart Vascular; circulatory system; Contrast Media; Contrast Agent; Contrast Drugs; Radiopaque Media; Data Analyses; Data Analysis; data interpretation; Developmental Biology; Drainage procedure; Drainage; Electronics; electronic device; Engineering; Environment; Feedback; Fluorescence; Future; Head; Hemoglobin; Housing; Illinois; image reconstruction; image construction; image generation; Institutes; Longitudinal Studies; long-term study; longitudinal outcome studies; longterm study; lymph nodes; Lymph Node Reticuloendothelial System; Lymph node proper; Lymphatic nodes; lymph gland; lymphnodes; Magnetic Resonance Imaging; MR Imaging; MR Tomography; MRI; MRIs; Medical Imaging, Magnetic Resonance / Nuclear Magnetic Resonance; NMR Imaging; NMR Tomography; Nuclear Magnetic Resonance Imaging; Zeugmatography; Maps; Methods; Nude Mice; Athymic Mice; Athymic Nude Mouse; Metastasis; Metastasize; Metastatic Lesion; Metastatic Mass; Metastatic Neoplasm; Metastatic Tumor; Secondary Neoplasm; Secondary Tumor; cancer metastasis; tumor cell metastasis; Neoplasm Metastasis; Neurosciences; optical; Optics; Patents; Legal patent; PET; PET Scan; PET imaging; PETSCAN; PETT; Positron Emission Tomography Medical Imaging; Positron Emission Tomography Scan; Rad.-PET; positron emission tomographic (PET) imaging; positron emission tomographic imaging; positron emitting tomography; Positron-Emission Tomography; Production; Common Rat Strains; Rat; Rats Mammals; Rattus; Research; Development and Research; R & D; R&D; research and development; Investigators; Researchers; Research Personnel; Research Resources; Resources; Robotics; Cell Communication and Signaling; Cell Signaling; Intracellular Communication and Signaling; Signal Transduction Systems; Signaling; biological signal transduction; Signal Transduction; Software; Computer software; Spatial Distribution; Specimen Processing; Specimen Handling; Medulla Spinalis; Spinal Cord; Progenitor Cells; stem cells; Tail; Technology; Testing; Time; tomography; single photon emission computed tomography; SPECT; SPECT imaging; Single-Photon Emission-Computed Radionuclide Tomography; Toxicology; Universities; Washington; Work; Generations; Imaging Techniques; Imaging Procedures; Imaging Technics; Drug Costs; Cost Savings; Bioluminescence; Organ; Label; improved; whole body imaging; whole body scanning; Peripheral; Area; Phase; Reporter Genes; Individual; Funding; Collaborations; Therapeutic; instrument; Photosensitiveness due to sun; sun sensitivity; Photosensitivity; programs; Scanning; subdermal; subcutaneous; Source; System; 3-D; 3D; three dimensional; 3-Dimensional; bioengineered tissue; engineered tissue; Tissue Engineering; experience; field based data; field learning; field test; field study; Performance; success; fluorophore; spinal cord regeneration; Animal Models and Related Studies; model of animal; model organism; Animal Model; novel; Modality; Modeling; drug development; animal facility; 3-D Imaging; 3D imaging; Three-Dimensional Imaging; drug discovery; Skin; image-based method; imaging method; imaging modality; Data; Detection; Imaging Instrument; Imaging Tool; Imaging Device; multi-modal imaging; multi-modality imaging; multimodality imaging; Multimodal Imaging; Research Contracts; Resolution; in vivo; Functional Imaging; Physiologic Imaging; physiological imaging; Laboratory Animal Production and Facilities; Research Animal Facility; Rodent Model; Small Business Innovation Research Grant; SBIR; Small Business Innovation Research; Validation; Characteristics; Molecular; Development; developmental; molecular imaging; molecule imaging; tissue regeneration; regenerate new tissue; regenerate tissue; regenerating damaged tissue; regenerating tissue; tissue regrowth; tissue renewal; tissue specific regeneration; Image; imaging; pre-clinical; preclinical; preclinical study; pre-clinical study; optical imaging; optic imaging; reconstruction; design; designing; nanoparticle; nano particle; nano-sized particle; nanosized particle; Three-dimensional analysis; 3-D analysis; 3-dimensional analysis; 3D analysis; Outcome; data acquisition; clinically significant; clinical significance; pre-clinical research; preclinical research; two-dimensional; 2-dimensional; human disease; mouse model; murine model; fluorescence imaging; fluorescent imaging; prototype; multimodality; multi-modality; Biological Markers; bio-markers; biologic marker; biomarker; animal model development; arm; photoacoustic imaging; optoacoustic imaging; in vivo imaging; imaging in vivo; animal imaging; imaging platform; data visualization; Metastatic breast cancer; imaging system; anatomic imaging; anatomical imaging; optoacoustic tomography; photoacoustic tomography; preclinical development; pre-clinical development; in vivo imaging system; IVIS SpectrumCT; IVIS imaging; IVIS optical imaging; IVIS spectral imaging; IVIS spectrum; IVIS system; in vivo optical imaging; Injections; imaging facilities; imaging center; human model; model of human; deep learning; three-dimensional visualization; 3-D visualization; 3-dimensional visualization; 3D visualization; regenerative biology; regeneration biology; stem cell self renewal; stem cell regeneration; ultrasound