Rebecca Richards-Kortum, PhD - US grants

This PYI award is to improve thebasic understanding and the application of optical spectroscopy to biomedical problems. Fluorescence and reflectance spectroscopy are powerful techniques for analyzing the chemical composition of tissues noninvasively. As a first step, the PI will analyze the information contained in a fluorescence spectrum from a multi-component, scattering tissue. Using imaging techniques, methods will be developed for characterizing the tissue as a function of position within the sample. It is expected that the proposed research will significantly enhance the treatment of diseases (such as atherosclerosis and neoplasia) where a quantitative understanding of tissue composition is essential.

This Presidential Faculty Fellow award will support studies to extract information from fluorescence and reflectance spectra of multi-component, scattering samples. Models of fluorescence will be developed and new methods to record fluorescence spectra and spectroscopic images will be investigated. This research has the potential to provide sensitive information about the chemical composition of tissue in a non-invasive way. This award also will support the teaching activities of the investigator. The activities include teaching an undergraduate physics course and developing demonstrations to illustrate the principles being taught. At the graduate level, the investigator will expand two courses in optics and continue to promote faculty involvement in a graduate research seminar.

confocal scanning microscopy; fiber optics; biomedical equipment development; bioimaging /biomedical imaging; human subject; clinical research;

DESCRIPTION (Adapted from Applicant's Abstract): The applicants proposed to identify, evaluate and improve the clinical role of optical spectroscopy and imaging for cervical pre-cancer detection. To achieve this goal, the following research projects will be conducted. First, a multi-pixel fluorescence imaging system that has been developed will be evaluated as a post-screening/pre-diagnostic filter for cervical pre-cancer detection. The second goal will be to incorporate diffuse reflectance spectra (measured using a flying spot scanner) from cervical tissues in vivo into our fluorescence based algorithms to determine if it can enhance their sensitivity and specificity for diagnostic applications. The specificity of the current fluorescence based clinical algorithms is limited in part by the presence of inflammatory tissues which appear to be diseased, spectroscopically. These algorithms are based on fluorescence spectra at 337, 380 and 460 nm excitation, which may not be optimum for discriminating between neoplastic and inflammatory tissues. The third objective will be to measure and compare the fluorescence spectra of cells that are predominant in inflammatory tissues to that of cultured neoplastic cervical epithelial cells at a series of excitation wavelengths over the ultraviolet and visible spectral regions. The applicants proposed to identify if there are optimal excitation wavelengths that are specific to inflammatory cells, and we will test these parameters in a future clinical investigation. The final component of our proposed research will be to explore the biochemical and morphological basis of the cervical tissue spectra measured in vivo. A Monte Carlo model of tissue fluorescence will be developed which will utilize the fluorescence spectra from cervical tissue microstructures and will also incorporate effects due to both absorption and scattering to describe the bulk tissue fluorescence.

This Integrative Graduate Education and Research Training (IGERT) award will support the establishment of a multidisciplinary graduate training program in Optical Bio-Molecular Engineering (OBE) that melds the more traditional areas of optical engineering, biological chemistry, and molecular biology. This activity is a joint effort of fourteen scientists and engineers from the Departments of Botany, Chemistry, Chemical Engineering, Electrical and Computer Engineering, Mechanical Engineering, Physics, and Zoology. This group of scientists and engineers has established research collaborations and has interacted extensively through the existing Graduate Programs in Molecular Biology and Biomedical Engineering at the University of Texas. Their collective expertise will provide the intellectual underpinning for the training of a diverse cadre of some seventy graduate students over the five-year tenure of the award. This IGERT activity in OBE draws upon recent advances in optical technologies that enable dynamic visualization and manipulation of processes within single cells and tissues with nanoscale resolution. Important features of the program include broad exposure of doctoral students to the field through research internships and seminars; interdisciplinary doctoral research guided by multiple mentors; and new courses that provide the multidisciplinary perspective necessary for the design and use of optical instrumentation for visualization and manipulation of tissues and cells. IGERT is a new, NSF-wide program intended to facilitate the establishment of innovative, research-based graduate programs that will train a diverse group of scientists and engineers to be well-prepared to take advantage of a broad spectrum of career options. IGERT provides doctoral institutions with an opportunity to develop new, well-focussed multidisciplinary graduate programs that transcend organizational boundaries and unite faculty from several departments or institutions to establish a highly interactive, collaborative environment for both training and research. In this first year of the program, support will be provided to seventeen institutions for new or nascent programs that collectively span all areas of science and engineering supported by NSF.

9872829
Richards-Kortum
The objective of this research is to develop new knowledge and technologies which integrate the advantages of optical coherence tomographic (OCT) imaging and optical spectroscopy. The aim is to enable high-contrast, molecular-specific optical imaging of early tumors in epithelial tissues. This will require the development of new biological models to allow rigorous testing of these concepts. Also, the knowledge gained from these models will be used to develop an optical system to address the clinically significant problem of ovarian cancer. The completion of these objectives will require: (1) fundamental advances in the understanding of the cellular basis of light scattering and how it can be manipulated with extrinsic agents, (2) technology development for simultaneous imaging and spectroscopy, and (3) development of molecular contrast agents and tools to test tools and developments.
***

There is an important need for improved screening and detection methods for cervical intra-epithelial neoplasia that are both sensitive and cost- effective. Recently, many groups have demonstrated that techniques based on quantitative optical spectroscopy have the potential to fulfill this need. Many optical techniques have shown promise for in vivo detection of pre-cancer including fluorescence spectroscopy, reflectance spectroscopy, multi-spectral fluorescence imaging, and multi-spectral reflectance imaging using polarized and unpolarized light. White it is clear than quantitative optical methods have the promise to deliver highly sensitive, specific and cost-effective screening and detection tools, the biological and morphological bases for the differences in optical spectra of normal and neoplastic cervic are not well understood. Moreover, the choice of illumination and detection wavelengths and geometries has been made arbitrarily on the basis of small in vitro studies. The specific goals of this proposal are five fold: (1) to conduct a series, , of studies to elucidate the biological and morphological basis for the difference between the fluorescence and reflectance spectra of normal and neoplastic cervical tissue in vivo, (2) to conduct a series of studies designed to explore how differences in the fluorescence and reflectance spectra of normal and neoplastic cervical tissues can be enhanced by using simple contrast agents such as acetic acid, toluidine blue, iodine and hyper- and hypo-tonic saline, (3) to develop analytic and computational models which describe tissue optical properties and measured spectra in terms of tissue biochemistry, morphology and architecture in the presence and absence of extrinsic contrast agents, (4) to use these models to predict which optical techniques, wavelengths, optical illumination and collection geometries and contrast agents will provide the best discrimination between normal and neoplastic cervical tissue, and (5) to develop inverse models which enable information about tissue biochemistry and architecture to be extracted from measured spectra to improve optical diagnostic algorithms for cervical pre-cancer. We believe that the proposed studies will yield important improvements in the sensitivity and cost-effectiveness of algorithms for detection of cervical pre-cancer based on optical spectra. In summary, the studies proposed here will develop a complete and quantitative understanding of the connections between tissue optical spectra and biochemistry, morphology and architecture.

0086736
Descour
The goal of this interdisciplinary research project is to develop miniature microscopes that can utilize the interaction of light with tissues in many modalities to image morphology and cytochemistry in vivo, yielding tools that provide better delineation of tumors. The investigators envision pen-sized, battery-powered multi-modal miniature microscopes (4Ms) designed to specifically image microscopic and molecular features of pre-cancer. The proposed miniature microscopes are being called multi-modal because of their potential for enabling different imaging modalities such as optical sectioning, 3-D spectral fluorescence imaging, and reflectance imaging. The size and cost of these microscopes will be small enough so that they can be used for wide-scale screening of disease. These tools will have broad applicability in many organ sites due to their very compact size and capability for imaging. The investigators will first apply these tools to improve detection of oral-cavity pre-cancers because of the oral cavity's easy accessibility. Furthermore, the assembled multidisciplinary expertise empowers the investigators to treat the design of miniaturized imaging sensors in an all-inclusive manner. In addition to developing 4M devices, the investigators will also design contrast agents specific for molecular alterations associated with pre-cancer that can be applied topically to significantly expand the imaging capability of miniature microscopes.

0119450
Richards-Kortum
This proposal brings together scientists from very diverse areas with the goal of developing new photonic probes and contrast agents for highly sensitive and selective detection of pre-cancers in vivo . Dr. Andrew Ellington will use the approaches of combinatorial chemistry to develop a library of aptamer molecules specific for biomolecular targets on the surface of cervical cancerous and pre-cancerous cells. He will use well-established cervical cell lines at different stages of cancer development provided by Dr. Lotan. Drs. Brian Korgel and Konstantin Sokolov will develop new photonic probes based on quantum dots (BK) and metal nanoparticles (KS). They will utilize both the aptamers developed by Dr. Ellington as well as well-known antibodies currently used in clinical histopathology. Dr. Rebecca Richards- Kortum will test the conjugates as molecular specific contrast agents using optical microscopy and spectroscopy. She will use cervical cancer cell lines provided by Dr. Lotan, three-dimensional tissue phantoms and fresh cervical tissue slices from Dr. Follen. Experiments with all three biological systems representing properties of normal and neoplastic cervix at different levels of complexity will be used to assess and refine the performance and detection scheme for the new contrast agents. This refinement will include preparing bioengineered aptamers with high affinity to cancer specific targets, tailoring optical properties of metal nanoparticles and quantum dots, optimizing conjugation procedures, and developing optimal imaging geometries.

[unreadable] DESCRIPTION (provided by applicant) Recent developments in photonic technology provide the ability to non-invasively image in vivo; these new cellular imaging technologies have the potential to dramatically improve the prevention, detection and therapy of epithelial cancers. In vivo confocal microscopy is a new technology that can provide detailed images of tissue architecture and cellular morphology in living tissue near real time. Pilot clinical trials show that confocal images of epithelial tissues can diagnose dysplasia with unprecedented specificity and sensitivity. Furthermore, flexible fiber optic instrumentation that permits high resolution in vivo imaging has been developed. In this renewal, a series of collaborative, integrated clinical and engineering studies are proposed to explore the clinical role of in vivo confocal imaging in the detection, management and therapy of caners of the oral cavity and to develop cost-effective approaches to provide clinical instrumentation for multi-center trials. The aims of this proposal are to: 1) Conduct a large prospective trial to estimate the sensitivity and specificity of reflectance based confocal imaging for detection of dysplasia in the oral cavity; 2) conduct a clinical trial to evaluate the role of in vivo reflectance confocal imaging to maximize surgical efficacy and minimize surgical morbidity by assisting in margin detection for tumor resection in the oral cavity; 3) conduct a clinical trial to examine the ability of in vivo reflectance confocal imaging to monitor response to induction chemotherapy in a group of young patients with cancer of the tongue; 4) assess the ability of in vivo reflectance confocal microscopy to provide a tool to monitor the field at risk without the need for biopsy, providing a powerful new intermediate endpoint marker to aid in chemoprevention studies. Finally, the miniaturized objectives designed in our previous work enable in vivo imaging but are expensive to manufacture and difficult to assemble. The fifth aim of this proposal is to design and construct inexpensive, injection-modeled optical elements that can be easily integrated with the inexpensive fiber optic imaging bundles used in current prototypes.

[unreadable] DESCRIPTION (provided by applicant): [unreadable] Worldwide, about 15% of human cancers are caused by mechanisms involving viral infection. In particular, specific types of human papillomavirus (HPV) are the principal cause of cervical cancer and its precursors. The goal of this application is to develop molecular specific contrast agents to enable low cost in vivo imaging of HPV induced cervical carcinogenesis. Most cases of cervical cancer occur in resource-poor settings, where there is insufficient infrastructure to carry out screening, diagnosis and treatment with conventional means. The molecular imaging contrast agents to be developed and tested in this application have the potential to provide sensitive and specific detection for cervical cancer and its precursors. We envision a topical delivery agent that can be applied to the cervix during a pelvic examination and interrogated using an inexpensive laser pointer (< $10 US), enabling combined screening, detection and therapy in a single visit, without the infrastructure and costs required to prepare and read Papanicolaou smears or biopsies. In the R21 phase of this application, we will show proof of concept of these contrast agents in cell and tissue culture model systems. With this reduction in risk, we will then proceed to the R33 phase of the application where a topical delivery system will be developed and pre-clinical trials will be conducted in human organ culture models and in an animal model. In the R21 phase, we will produce three different contrast agents to selectively label cells expressing high levels of two oncoproteins associated with HPV 16 induced cervical carcinogenesis: E6, and E7 and one protein associated with abnormal expression of the cell cycle, p16. Commercially available monoclonal antibodies will be conjugated to gold nanoparticles. One contrast agent will be validated in cell culture and tissue culture models. Successful completion of these experiments will demonstrate the potential for in vivo molecular imaging of a key step in cervical carcinogenesis produced by HPV infection. These pathfinder experiments have been designed to test whether specific labeling can produce strong optical signatures in model systems of progressively increasing biologic complexity. In the R33 phase of the application, these goals are extended to include the next steps required for successful clinical implementation, including a topical delivery system and pre-clinical testing in human organ culture models and an animal model. [unreadable] [unreadable]

The advent of cellular and molecular-based detection and therapeutic technologies has begun to transform the way we think about human disease. Instead of relying only on clinical evaluation and general physiologic markers to detect disease and select therapy, physicians are beginning to use cellular and molecular biomarkers of disease to select and even to design the optimized therapeutic regime for an individual patient. To achieve the promise of this approach requires tools to detect molecular and cellular markers of disease in vivo and to monitor their modulation in response to therapy. Cellular and molecular imaging have the ability to dynamically visualize biomarkers of disease in single living cells and tissues with microscopic resolution, yielding a fundamental change in the way we diagnose disease, and select and monitor therapy. To achieve the promise of cellular and molecular imaging requires engineers and scientists with a wide breadth of skills in imaging science, contrast agent development, and design of tools to monitor cellular and molecular-based therapeutics.

The goal of this IGERT project is to develop an interdisciplinary graduate training program focused on cellular and molecular imaging for diagnostics and therapeutics. This interdisciplinary program builds on our existing IGERT grant in Optical Molecular Bio-Engineering. Over the last four and a half years, our team has trained students to develop and use new photonic technologies, adaptable and specific contrast agents, and computers to address both fundamental biological questions and the need for improved diagnostic imaging modalities. At the same time, the University of Texas at Austin, the UT M.D. Anderson Cancer Center and the UT Health Science Center at Houston have invested heavily in new interdisciplinary programs in Biomedical Engineering. In this competitive renewal, we integrate these efforts, to develop a multidisciplinary training program in cellular and molecular imaging for diagnostics and therapeutics.

The clinical use of cellular and molecular imaging hinges on the availability of a cadre of professionals with inter-disciplinary training spanning imaging science, biomarkers of disease, design and use of molecular contrast agents, and the principles of cellular and molecular based therapies. This project will develop an inter-disciplinary new pathway, which synthesizes these fields in a single graduate degree. The degree program will incorporate four important inter-disciplinary features. (1) A doctoral portfolio program of coursework will prepare students to carry out inter-disciplinary research in this field. (2) Students will carry out inter-disciplinary research under a new advisory structure, where co-advisors from different disciplines supervise the student's research. The thesis committee will have broad participation from faculty in the Colleges of Engineering and Natural Sciences as well a clinical mentor. (3) Students will receive training in technology assessment and transfer, to help them appreciate the spectrum of translational research as it leads to commercial products. (4) Students will participate in at least one internship where they see the application of their research in a different setting: Clinical internships allow students to participate in translational, clinical research; industrial internships enable students to participate in technology development; and international internships enable them to carry out research and development with a global perspective.

The intellectual merit and broader impacts of this IGERT project reside in the integration of inter-disciplinary research and education that will bring together teams of students, faculty and clinicians with diverse skill sets to develop and evaluate novel molecular and cellular imaging systems for diagnostics and therapeutics. These teams will collaborate to develop and present courses and seminars, to define important clinical problems and to develop and evaluate engineering based solutions. Through participation with historically minority Texas undergraduate institutions, we will recruit graduate students from underrepresesented groups to participate in this program. As we focus on the parallel development, assessment, and transfer of technology, we will create key partnerships with industry. These partnerships will provide our students with information about a variety of career opportunities, and will expedite the process of bringing the research advances developed here to general medical practice.

IGERT is an NSF-wide program intended to meet the challenges of educating U.S. Ph.D. scientists and engineers with the interdisciplinary background, deep knowledge in a chosen discipline, and the technical, professional, and personal skills needed for the career demands of the future. The program is intended to catalyze a cultural change in graduate education by establishing innovative new models for graduate education and training in a fertile environment for collaborative research that transcends traditional disciplinary boundaries. In this sixth year of the program, awards are being made to institutions for programs that collectively span the areas of science and engineering supported by NSF.

DESCRIPTION (provided by applicant): Cancer is a major public health problem. Currently, classification of cancer is based on phenotypic markers. The identification of unique molecular markers of cancer has led to development of new molecular cancer therapies. Movement toward a molecular characterization of cancer would have important clinical benefits, including (1) detecting cancer earlier, (2) predicting risk of precancerous lesion progression, (3) detecting margins in the operating room in real time, (4) selecting molecular therapy rationally and (5) monitoring response to therapy in rear time at a molecular level. Imaging the molecular features of cancer requires molecular-specific contrast agents which can safely be used in vivo as well as cost-effective imaging systems to rapidly and non-invasively image the uptake, distribution and binding of these agents in vivo. Radiographic imaging modalities such as CT and MRI, although useful for delineating the deep extent of advanced carcinomas, are not sufficiently sensitive to detect small, intraepithelial lesions. Optical imaging is a new modality which enables real time, high resolution imaging of epithelial tissue. Optical imaging systems are inexpensive, robust and portable. Optical imaging systems are ideally suited for early detection of intraepithelial disease and to assess tumor margins and response to therapy. The goal of this proposal is to integrate development of optical imaging systems and contrast agents with advances in functional genomics. We will develop molecular-specific, optically active contrast agents that can be applied topically. We will also develop inexpensive, rugged and portable imaging systems to monitor the three-dimensional profile of targeted biomarkers. These contrast agents and imaging systems will have broad applicability to many types of cancer; here, we will develop and test agents and imaging systems for the cervix, oral cavity and the lung, which represent more than 20% of both tumor incidence and mortality worldwide. We will test the safety and efficacy of these contrast agents and imaging systems in animal models, providing data to support phase I and II clinical trials. The aims of this proposal are to: (1) Develop optically active contrast agents to target four molecular signatures of neoplasia, including EGFR, MMP, telomerase and alpha v integrin; (2) to identify promising new biomarkers for which contrast agents will be developed using SAGE libraries, and to identify promising molecular probes for novel contrast agents using combinatorial methods; (3) to develop inexpensive, portable optical systems to image the morphologic and molecular signatures of neoplasia noninvasively in real time; and (4) to test these agents, delivery formulations and imaging systems in living biological systems of progressively increasing complexity. (5) Our final aim is to integrate these studies to develop a miniature imaging system, which when coupled with the contrast agents developed here, can be used for real time, molecular detection of neoplasia and to monitor, at the molecular level, whether a lesion is responding to therapy.

The goal of this project is to transform the Schools of Science and Engineering at Rice University by increasing the number of women faculty; strengthening the gender-neutrality of the climate in a way that identifies and values the unique skills of each individual and rewards contributions; and enhancing opportunities for women to assume and succeed in leadership positions. Approximately one-third of the faculty in science, mathematics, engineering, and technology areas at Rice will reach normal retirement age within the next 8 years. This should provide an exceptional opportunity to seek to significantly increase the numbers of women faculty, including women of color. The ADVANCE program will capitalize on the strong commitment to gender equity at Rice University and robust linkages to the leadership of the institution to address the key issues of recruitment, retention, and climate that affect women in academia. To achieve this transformation, three specific goals are being pursued:

1. Increase the number of women at Rice
Aspiration: Hiring and retention of women faculty at levels that better reflect available pools of Ph.D.s within disciplines.
Recruitment Activities: Workshop for Career Success, database of qualified applicants, introducing best recruitment practices, engaging non-traditional entry tracks for women to enter the academy.
Retention Activities: Workshop for Faculty Success, database of service and teaching, introducing best practices for chairs and staff.
2. Create a positive and welcoming work environment for women at Rice
Aspiration: Women and men report similar levels of satisfaction in climate surveys.
Individual Activities: Mentoring, coaching, and reverse mentoring.
Institutional Activities: Engagement of department chairs in institutional transformation, updating institutional policies to increase opportunities for success of women faculty, data collection and analysis, and discourse on leadership.
3. Evaluate what works to advance women and share this information
Aspiration: Publications in professional journals and materials made readily available.
Evaluation: Critical assessment by social scientists to produce publishable data as well as general guidelines for effective action.
Dissemination: Use of website and Connexions as mechanisms to provide easy access to materials for adaptation by other institutions and holding a national Workshop on Best Practices.

The success of this program provides the opportunity to distinguish Rice for its excellent climate and for its efforts in highlighting and encouraging achievement of women faculty with model activities that can be adapted and applied to other institutions. The series of initiatives focused on dissemination of developed materials enables project knowledge to be broadly available to other institutions undertaking transformational initiatives.

[unreadable] DESCRIPTION (provided by applicant): [unreadable] A new short course in "Translational Optical Molecular Imaging: Nano to Macro" will be hosted by three neighboring institutions, Rice University, The University of Texas M.D. Anderson Center, and Baylor College of Medicine. Our strategy is to bring together leaders in this field to discuss paths that lead to their advancements and to plot new paths for the future in the area of optical imaging for clinical applications. Our course faculty are a world-renowned list of researchers with about half coming from the Texas Medical Center and half from leading institutions such as Stanford, MIT, Johns Hopkins, [unreadable] Harvard, Georgia Tech, etc. In addition to the lectures that will be presented by our invited course faculty, a unique feature of this course is the ability to interact with leading companies invested in the future of optical imaging strategies. These include Zeiss, Olympus, Molecular Probes/Qdots, General Electric, Phillips, Siemens, etc. The ability of research scientists to interact with leading companies will both facilitate collaborative interactions and guide the focus of corporate research and development programs, playing a large role in where efforts are placed to achieve the greatest advancements. [unreadable] The purpose of this proposal to request funds to support young investigators and graduate students and postdoctoral fellows to attend the short course. The objectives of this short course on "Translational Optical Molecular Imaging: Nano to Macro" are as follows: [unreadable] 1. To educate trainees and faculty about state-of-the-art optical imaging strategies for basic research and clinical practice and facilitate career development. [unreadable] 2. To further the development of novel optical imaging technologies that can be used for disease detection and treatment. [unreadable] 3. To establish and strengthen productive relationships between basic and clinical labs within the Texas Medical Center. [unreadable] 4. To establish and strengthen relationships with industry partners for the development of new technologies. [unreadable] [unreadable] [unreadable]

DESCRIPTION (provided by applicant): Recent advances in confocal microscopy have demonstrated the potential of optical imaging to provide molecular images with sub-cellular resolution. In this proposal, we extend our work in high resolution confocal microscopy, based on a much simpler alternative, based on our observation that high resolution optical images of the top 1-2 layers of cells can be obtained without the need for a confocal imaging gate if the cells are placed in direct contact with a flat optical window. Here we propose a new class of optical imaging device - the integrated optical needle (ION) - which can be inserted into a small gauge needle and advanced through tissue to acquire images of tissue at the distal tip of the needle with sub-cellular resolution in real time;the use of targeted contrast agents or vital stains can yield additional contrast as well as functional and/or molecular imaging capability. The first version of the optical needle, described in Aim 1, is based on a flexible, coherent fiber-optic bundle coupled to macroscopic imaging optics and a CCD image sensor. The under-sampling associated with the structure of the fiber bundle used in Aim 1 limits the spatial resolution. The second approach, an integrated optical needle (ION) described in Aims 2 and 3, uses LIGA technology to integrate miniature lenses fabricated using grayscale lithography, together with the light source and image sensor to yield a miniature microscope which can be inserted through a needle. This ION consists of a NA = 0.4 microscope that can image with 1 micron lateral resolution from a 250 um field of view;it is designed to fit within a needle that has an inner diameter of 1 mm. We will carry out a variety of preclinical tests using cell culture, tissue culture, animal models and pilot clinical trials to evaluate the relative performance of these systems. The question to be addressed is whether imaging without a pinhole or other depth-sectioning technique yields images of sufficient quality to appreciate cellular detail required clinically, and to determine whether out-of-focus and scattered photons reduce contrast to too large a degree. Our preliminary results indicate that it is possible to obtain good images of cellular detail in human subjects using contrast agents in routine clinical use. However, to judge image quality with the various devices and contrast agents, we will use quantitative measures of image resolution, image contrast and image SNR. Research proposed here will develop a new tool for image guided tissue assessment in real time, and confirmatory biopsy at image directed locations. The ION can image a broad range of contrast agents enabling molecular information from specific sites. The cost of the integrated optical needle will be low even in small quantities (below $2,300) but there is great potential to further reduce this cost when manufactured in quantity.

DESCRIPTION (provided by applicant): Our goal is to develop an integrated diagnostic test for HIV-1 viral load with high sensitivity, specificity, reliability, and reproducibility for use in minimal infrastructure settings. Such a test, applied in perinatal HIV-1 diagnosis, could save 180,000 DALYs each year if 5% of the targeted population has early access to therapy, and up to 2.5 million DALYs each year if 100% of the population has access to treatment [2]. It would also help to overcome one of the major challenges to universal access to therapy: the lack of adequate diagnosis and treatment of pediatric HIV-1 disease (WHO) [1]. Currently the most sensitive and reliable assays to quantify HIV-1 viral load rely on nucleic acid amplification and detection, but these tests often require sophisticated instrumentation and expensive reagents. Alternatively, the high analytical sensitivity of oligonucleotide-coated gold nanoparticles as targeting/reporting agents in nucleic acid tests has been demonstrated [2, 3]. To achieve the required sensitivity, we will develop an HIV-1 viral load assay which integrates isothermal PCR amplification of HIV-1 RNA with the use of targeted gold nanoparticles and colorimetric quantification of test results. We will develop this assay for use in low - resource settings, with minimal infrastructure requirements and a sensitivity and specificity comparable to that of commercial viral load assays available in the developed world. The specific aims of the proposal are to: (1) Develop an inexpensive, sensitive, and specific diagnostic test for determining type 1 HIV viral loads of seropositive patients in low-resource settings. The assay will combine: target isolation and isothermal amplification technologies to yield at least 104 fold amplification from samples containing a minimum of 1000 viral copies/ml. An oligonucleotide-targeted gold nanoparticle detection assay and a quantitative read-out will be then used to detect to achieve a dynamic detection range from 103 to 106 HIV-1 viral copies/ml at the POC. (2) Validate the performance of this assay for HIV-1 viral load determination in clinical specimens. In collaboration with Dr. Richard Sutton from Yale School of Medicine who has expertise in molecular biology of HIV-1, we will test the ability of the assay to detect RNA from whole viral particles, and from group M clades on an NIH/UNAIDS reference panel, comparing the assay to RT-PCR. In collaboration with Dr. Elizabeth Molyneux from Queen Elizabeth Central Hospital, Blantyre, Malawi who has expertise in clinical diagnosis of HIV-1, we will carry out a pilot study to determine the sensitivity and specificity of this new assay. Personnel from Dr. Molyneux's team will travel to Houston to learn the assay, and members of the Richards-Kortum lab will travel to Malawi to work with her group to evaluate the assay in pediatric clinical samples relative to the gold standard of dried blood spot RT-PCR. PUBLIC HEALTH RELEVANCE: Our goal is to develop an integrated diagnostic test for HIV viral load with high sensitivity, specificity, reliability, and reproducibility for use in minimal infrastructure settings. Viral load determination is needed to determine when to initiate therapy, monitor compliance, and most importantly, as an early indicator of therapeutic failure. Despite the encouraging trends in the scale-up of antiretroviral treatment in low- and middle-income countries reported by the WHO, reliable and accurate HIV load testing has yet to be introduced into the management of infected patients in low resource settings and remains one of the major challenges to universal access to therapy [1]. Such a diagnostic test, applied in perinatal diagnosis, could save 180,000 DALYs each year if 5% of the targeted population has early access to therapy, and up to 2.5 million DALYs if 100% of the population has access to treatment [2].

DESCRIPTION (provided by applicant): Esophageal adenocarcinoma (EAC) has one of the fastest rising rates of incidence in the US. Unfortunately, the five-year-survival for patients diagnosed with EAC is only 10%. EAC develops primarily in patients with Barrett's esophagus (BE). Endoscopic screening and biopsy is recommended for at-risk individuals. However, standard white-light endoscopic examination frequently misses areas of early neoplasia, which are often clinically indistinguishable from normal mucosa and/or inflammatory changes. Studies have shown that as many as 43-57% of early cancers can be missed by this method. Thus, there is an important need for new endoscopic technologies which improve the ability of clinicians to identify precancerous lesions and early cancers with high sensitivity and specificity. The goal of this proposal is to develop, optimize and validate novel multi-modal, multi-scale optical imaging platforms for non-invasive, early detection of esophageal neoplasia based on optical imaging. We will collaborate with colleagues at Pentax, Inc. to design and test multi-modal endoscopic imaging systems for early detection of neoplasia in Barrett's esophagus. Widefield endoscopic imaging will be used initially to screen the surface area at risk to identify abnormal sites with high sensitivity;suspicious areas will then be imaged with much higher spatial resolution to achieve high diagnostic specificity. Both wide field and high resolution technologies will be integrated into a single endoscopic platform to increase the ease and accuracy of endoscopic cancer screening and surveillance. In sequential clinical studies, we will first separately optimize the performance of wide field endoscopic imaging and high resolution imaging. We will then integrate the wide field and high resolution imaging systems and validate their accuracy for the detection of neoplasia in subjects with Barrett's esophagus, the precursor to esophageal adenocarcinoma. Lastly, we will develop an image atlas of typical wide-field and high-resolution images, interpretation criteria, and histopathology to train future users and serve as an educational resource. PUBLIC HEALTH RELEVANCE: Improved identification and treatment of early cancers and precancerous lesions represents the most important opportunity to reduce the morbidity and mortality of cancer. This is particularly true for esophageal adenocarcinoma, a malignancy with a rapidly rising incidence rate and uniformly poor survival, primarily due to diagnosis at a late, incurable stage.

DESCRIPTION (provided by applicant): This research will be done primarily in Botswana at the University of Botswana/Princess Marina Hospital in collaboration with Doreen Ramogola-Masire, as an extension of NIH Grant No. R01EB007594, 7-1-2008 to 6-30-2012. In 2007, cervical cancer accounted for over 24% of all cancers among women in Botswana. With studies establishing HIV as a cofactor in the development of HPV and cervical neoplasia and the incidence of HIV/AIDS in the country continuing to rise, early diagnosis of cervical cancer and its precursors is a key priority. This proposal integrates a series of collaborative studies to develop, test, and implement optical imaging technologies to improve cervical cancer screening in Botswana. In Aim 1, we will jointly develop a wide-field imaging system designed specifically to image the cervix. We will combine this system with a high-resolution microendoscope developed under the parent NIH grant to form a multimodal imaging platform. Members of Dr. Richards-Kortum's research group will travel to work with local biomedical engineers at the Princess Marina Hospital to assemble the imaging platform in Botswana, establishing the necessary infrastructure and expertise to build, test, and maintain optical imaging systems on-site. In Aim 2, Dr. Doreen Ramogola-Masire, an obstetrician gynecologist with extensive experience establishing and managing cancer screening programs in Africa, will lead a pilot clinical study to test these systems in Botswana, collecting wide-field and high-resolution image data in reflectance and fluorescence modes. In Aim 3, we will work together to analyze the clinical data to establish the imaging mode, or combination of modes which demonstrate the highest degree of correlation with disease state. This information will be used to generate image overlay maps to visually indicate the probability of disease, aimed at assisting the non-specialist healthcare provider in clinical decision making. Our long-term hypothesis is that quantitative, digital wide-field imaging combined with high-resolution cellular imaging will assist in the early detection of cervical cancer through improved screening in developing countries such as Botswana. PUBLIC HEALTH RELEVANCE: Cervical cancer is the leading cause of cancer death for women in developing countries. Optical imaging technologies can potentially assist healthcare providers in low-resource settings to improve the accuracy and coverage of screening programs. This proposal aims to develop and test wide-field and high-resolution optical imaging systems in a pilot clinical study in Botswana.

DESCRIPTION (provided by applicant): Esophageal adenocarcinoma (EAC) has one of the fastest rising rates of incidence in the US. Unfortunately, the five-year-survival for patients diagnosed with EAC is only 10%. EAC develops primarily in patients with Barrett's esophagus (BE). Endoscopic screening and biopsy is recommended for at-risk individuals. However, standard white-light endoscopic examination frequently misses areas of early neoplasia, which are often clinically indistinguishable from normal mucosa and/or inflammatory changes. Studies have shown that as many as 43-57% of early cancers can be missed by this method. Thus, there is an important need for new endoscopic technologies which improve the ability of clinicians to identify precancerous lesions and early cancers with high sensitivity and specificity. The goal of this proposal is to develop, optimize and validate novel multi-modal, multi-scale optical imaging platforms for non-invasive, early detection of esophageal neoplasia based on optical imaging. We will collaborate with colleagues at Pentax, Inc. to design and test multi-modal endoscopic imaging systems for early detection of neoplasia in Barrett's esophagus. Widefield endoscopic imaging will be used initially to screen the surface area at risk to identify abnormal sites with high sensitivity; suspicious areas will then be imaged with much higher spatial resolution to achieve high diagnostic specificity. Both wide field and high resolution technologies will be integrated into a single endoscopic platform to increase the ease and accuracy of endoscopic cancer screening and surveillance. In sequential clinical studies, we will first separately optimize the performance of wide field endoscopic imaging and high resolution imaging. We will then integrate the wide field and high resolution imaging systems and validate their accuracy for the detection of neoplasia in subjects with Barrett's esophagus, the precursor to esophageal adenocarcinoma. Lastly, we will develop an image atlas of typical wide-field and high-resolution images, interpretation criteria, and histopathology to train future users and serve as an educational resource.

DESCRIPTION (provided by applicant): Oral cancer is the 6th most common cancer worldwide. Despite the easy accessibility of the oral cavity for screening, oral cancer has one of the lowest 5-year survival rates of all cancers. Oral cancer is thought to arise as a result of fied cancerization, where, often in response to tobacco and alcohol exposure, wide areas of the mucosal surface develop subclinical carcinogenetic changes. The poor outcomes of oral cancer arise primarily because: (1) most patients are diagnosed at a late stage since the molecular changes that put patients at risk of neoplasia often do not give rise to clinically visible lesions and (2) a large fraction of patients treated for oral cancer develop subsequent cancers because areas of field cancerization persist following treatment and are not clinically visible. The development and progression of oral cancer is ultimately a molecular process, reflecting a complex succession of genetic changes within the field-at-risk. Ultimately tumor-initiating stem cells give rise to aggressive clones within a mucosal field-at-risk, resulting in malignant progression. While much progress has been made to understand the molecular alterations associated with oral cancer progression, this research has not yet led to improvements in early detection mainly because molecular analysis methods are costly and can only be carried out with tissues obtained from invasive biopsies. There is increasing evidence to suggest that key molecular alterations result in phenotypic changes that can be measured clinically at the point-of-care. Recent studies by our group and others suggest that multi-modal optical imaging can image changes in tissue fluorescence and nuclear morphometry to identify high grade oral precancer and early cancer with significantly improved sensitivity and specificity compared to visual examination; moreover, changes in optical properties correlate strongly with molecular markers associated with neoplastic progression. The goal of this proposal is to validate the ability of multimodal optical imaging to improve early detection and to determine whether risk-related optical markers (RROMs) can be used to predict the likelihood of malignant progression. We will perform longitudinal studies in patients with oral lesions using cutting edge autofluorescence and microendoscopy technology with automated diagnostic algorithms. In an animal model of oral cancer, we will combine optical imaging and novel tissue preparation techniques, which render tissue optically transparent and macromolecular permeable, to assess the temporal and spatial correlations of molecular alterations to phenotypic changes during development and progression of oral cancer. With this data, we propose to develop and validate predictive models relating RROMs to malignant transformation.

DESCRIPTION (provided by applicant): Cervical cancer was previously the leading cause of cancer-related death among women in the US; however, incidence and mortality have decreased by >70% due to the introduction of screening programs to detect early cervical cancer and its precursors. In stark contrast, cervical cancer continues to be the 1st or 2nd leading cause of cancer death among women in low- and middle-income countries (LMICs). There is a significant need for appropriate cervical cancer screening and diagnostic tools that can be used in resource- limited settings. The World Health Organization's recommendations for cervical cancer prevention methods in low-resource settings focus on screen-and-treat strategies using visual inspection with acetic acid (VIA) or HPV testing, alone or in combination. However in many large studies, the positive predictive value of VIA and HPV testing have been found to be <10%. Thus, based on VIA or HPV screening alone, >90% of women with a positive test result would be inappropriately treated in a see-and-treat setting. Here, we propose to optimize and validate a high resolution microendoscope (HRME) to be used in see- and-treat programs to improve specificity without reducing sensitivity. The goal of this application is to optimize and validate the performance of the HRME for real-time diagnosis of cervical cancer in urban and rural settings in Brazil. In the UH2 phase, we will demonstrate successful implementation of the HRME in a novel mobile diagnostic and treatment unit for real-time diagnosis and treatment of cervical precancer in screen-positive women in a single visit in order to reduce the number of women lost to follow-up. In the UH3 phase, we will carry out a study involving over 12,000 women to validate that the HRME has improved diagnostic sensitivity and specificity compared to VIA and colposcopy for combined diagnosis and treatment in a single visit. Our project will validate the use of HRME as a real-time, in vivo diagnostic tool that can b applied in a variety of scenarios as an alternative to the traditional diagnostic methods of colposcopy and biopsy, which are impractical in many LMICs. Our team includes expertise in bioengineering (Rice University) and cervical cancer prevention and treatment (UT MD Anderson Cancer Center and Barretos Cancer Hospital). Our teams have worked together for two years to develop affordable, effective technologies for cervical cancer prevention in low-resource settings. To ensure that results of this work lead to sustainable implementation and scale-up within and beyond Brazil, we are teamed with the Global Coalition Against Cervical Cancer, an NGO that assists LMICs in the implementation of comprehensive, sustainable, and effective cervical cancer prevention and control. Our development partner is Becton Dickinson, a multi-national medical technology company with a strong commitment to global efforts to improve women's health.

DESCRIPTION (provided by applicant): Cervical cancer continues to be the 1st or 2nd leading cause of cancer death among women in low- and middle- income countries. In the U.S., low socioeconomic status (SES) is strongly correlated with poor cervical cancer survival. In resource-limited settings in the U.S. and abroad, there is a significant need for new point-of-care diagnostics that enable combined detection and treatment of cervical precancer in a single visit. To date, global attempts to implement see & treat protocols have been limited by the extremely low specificity of the three existing diagnostic approaches that can be used at the point-of-care. See & treat protocols based on imaging (colposcopy or visual inspection with acetic acid (VIA)) or biomarker detection (HPV DNA testing) result in high rates of overtreatment, subjecting patients to unnecessary procedures and wasting healthcare resources. We hypothesize that a combination of imaging and biomarker detection can reduce the high false positive rate of current cervical screening tools. Increased nuclear-to-cytoplasmic (N/C) ratio is one of the best known phenotypic biomarkers of cervical precancer, but can currently only be assessed from cytology or biopsy. We have developed an investigational imaging method, high-resolution microendoscopy (HRME), to identify cervical lesions in situ with increased N/C ratio. In Aim 1, we will adapt the HRME to a mobile platform which incorporates a cell phone to capture, display, analyze, and transmit images. We will then evaluate the performance of the mobile HRME (mHRME) with an existing biomarker (HPV DNA testing) and an investigational biomarker (HPV E7 oncoprotein). In Aim 2, we will evaluate performance of the mHRME in two globally relevant contexts: (A) in a clinical evaluation of 429 women in the U.S., we will assess whether addition of mHRME imaging improves specificity of colposcopy, without significantly reducing sensitivity; (B) in a clinical evaluation of 4,592 women in El Salvador, we will assess whether mHRME imaging improves the specificity of screening by VIA alone, HPV DNA testing alone, or with HPV DNA testing followed by VIA. Overexpression of HPV E7 oncoprotein is one of the best known molecular biomarkers of cervical precancer. To further improve specificity of molecular testing, in Aim 3 we will develop a rapid, low-cost, laterl flow test to detect E7. We will assess test performance using cervical specimens in a nested case-control of 270 patients with and without cervical precancer who participated in Aim 2. This work will provide clinicians in the U.S. and globally with robust, affordable, integrated, point-of care tools to directly image phenotypic changes and detect molecular markers associated with the development and progression of cervical precancer, addressing the poor specificity of current methods. Together, these tools will improve the efficacy and cost-effectiveness of early detection of cervical precancer, allowing diagnosis and treatment in a single visit to prevent the development of invasive cervical cancer.

? DESCRIPTION (provided by applicant): In the last decade there has been an exponential growth in nanotechnology research in cancer demonstrating that nanotechnology could provide unique and otherwise unattainable solutions to cancer management including very early cancer detection, accurate molecular specific diagnosis and treatment that diminishes side effects. However, achieving this promise is extremely challenging because it requires overcoming multiple constraints imposed by translational barriers in clinical applications of nanomaterials that is multiplied by complexity of cancer biology. Currently, there is a growing gap between new discoveries coming at a fast pace from academic labs and their translation into clinic. Therefore, there is an urgent need in addressing this gap in cancer nanotechnology translational pipeline. To this end, we have designed a novel training program to educate future leaders in the broad field of nanotechnology with specific interests in cancer-related applications, who are keenly aware of the needs and demands of clinical environment as well as of major challenges of translational research. We believe that the only way to train cancer translation minded Ph.D. researchers is to insert them into the environment of an outstanding cancer center. Therefore, our program is based on a close collaboration between The University of Texas MD Anderson Cancer Center and Rice University. As part of our program, we have developed a comprehensive plan for recruiting trainees from underrepresented minority groups that are historically underrepresented in health-related research, including women, minority individuals, and individuals with disabilities. Our training program includes multidisciplinary mentorship of translational research projects combined with multidisciplinary, hands-on coursework and seminar experiences. All trainees will work with at least two program faculty mentors (one from Rice and one from MD Anderson) to define and carry out an independent research problem. Didactic coursework will prepare them to contribute to research projects that directly address barriers to translation of nanotechnology-based approaches and to develop the skills needed to define and lead such projects. Incoming trainees will participate in a unique 2-week-long boot camp in Cancer Management and Nanotechnology that provides an overview of current opportunities and barriers in the field. Trainees will develop foundational background in the field by taking four courses related to translational cancer or nanotechnology topics. Trainees will gain an appreciation for federal resources to assist in cancer nanotechnology research by taking a trip to the NCI Nanotechnology Characterization Lab. Finally, trainees will gain important lab management skills by participating in a short hands-on course providing an introduction to laboratory and project management. At the end of the program, fellows will have a deep understanding of translational research in cancer nanotechnology, with the most important component being the demonstrated ability to carry out independent translational research in this challenging multidisciplinary field.

DESCRIPTION (provided by applicant): Cervical cancer continues to be the 1st or 2nd leading cause of cancer death among women in low- and middle- income countries. In the U.S., low socioeconomic status (SES) is strongly correlated with poor cervical cancer survival. In resource-limited settings in the U.S. and abroad, there is a significant need for new point-of-care diagnostics that enable combined detection and treatment of cervical precancer in a single visit. To date, global attempts to implement see & treat protocols have been limited by the extremely low specificity of the three existing diagnostic approaches that can be used at the point-of-care. See & treat protocols based on imaging (colposcopy or visual inspection with acetic acid (VIA)) or biomarker detection (HPV DNA testing) result in high rates of overtreatment, subjecting patients to unnecessary procedures and wasting healthcare resources. We hypothesize that a combination of imaging and biomarker detection can reduce the high false positive rate of current cervical screening tools. Increased nuclear-to-cytoplasmic (N/C) ratio is one of the best known phenotypic biomarkers of cervical precancer, but can currently only be assessed from cytology or biopsy. We have developed an investigational imaging method, high-resolution microendoscopy (HRME), to identify cervical lesions in situ with increased N/C ratio. In Aim 1, we will adapt the HRME to a mobile platform which incorporates a cell phone to capture, display, analyze, and transmit images. We will then evaluate the performance of the mobile HRME (mHRME) with an existing biomarker (HPV DNA testing) and an investigational biomarker (HPV E7 oncoprotein). In Aim 2, we will evaluate performance of the mHRME in two globally relevant contexts: (A) in a clinical evaluation of 429 women in the U.S., we will assess whether addition of mHRME imaging improves specificity of colposcopy, without significantly reducing sensitivity; (B) in a clinical evaluation of 4,592 women in El Salvador, we will assess whether mHRME imaging improves the specificity of screening by VIA alone, HPV DNA testing alone, or with HPV DNA testing followed by VIA. Overexpression of HPV E7 oncoprotein is one of the best known molecular biomarkers of cervical precancer. To further improve specificity of molecular testing, in Aim 3 we will develop a rapid, low-cost, laterl flow test to detect E7. We will assess test performance using cervical specimens in a nested case-control of 270 patients with and without cervical precancer who participated in Aim 2. This work will provide clinicians in the U.S. and globally with robust, affordable, integrated, point-of care tools to directly image phenotypic changes and detect molecular markers associated with the development and progression of cervical precancer, addressing the poor specificity of current methods. Together, these tools will improve the efficacy and cost-effectiveness of early detection of cervical precancer, allowing diagnosis and treatment in a single visit to prevent the development of invasive cervical cancer.

Abstract: Esophageal squamous cell neoplasia (ESCN) is the sixth leading cause of cancer death worldwide with only 1 in 5 patients surviving three or more years due to diagnosis at an advanced stage. When detected early, endoscopic treatment can be performed resulting in dramatically improved survival (>90%). Unfortunately, early neoplasia is difficult to visualize on routine white light endoscopy. Pre-cancerous lesions and carcinoma often appear as small erosions, flat lesions, or normal mucosa and recognition of neoplasia is difficult even for experienced endoscopists. Currently, endoscopic screening is performed in high-risk patients using Lugol's chromoendoscopy (LCE). Although LCE increases sensitivity to >95%, specificity remains <65%, resulting in many unnecessary biopsies and increased cost. Recent studies have shown that Narrow-Band Imaging with magnification has significantly higher positive predictive value compared to LCE. However, it has proven difficult to implement any esophageal screening programs in low-resource settings due to the high cost of conventional endoscopy equipment (>$40,000), the need to sedate patients undergoing conventional endoscopy, and the lack of infrastructure to process equipment and biopsies. Capsule endoscopy is an attractive alternative for low-resource settings because patients do not require sedation. However, current capsule endoscopy systems lack sensitivity to diagnose ESCN because of limited spatial resolution; moreover, commercially available capsule endoscopy systems are expensive ($25,000). The goal of this proposal is to develop a $2,500 high-resolution capsule endoscopy system to allow less experienced providers to screen for ESCN in low resource settings. The system has two components: a reusable, $50 tethered capsule that is swallowed to collect images of the mucosa and supporting hardware to manipulate and display the collected data on a tablet computer. The system is named ScanCap because the tether causes the capsule to rotate as it descends through the esophagus during peristalsis so that it captures high resolution images from the entire esophagus. The system is portable, battery operated and designed for use by non-physicians in underserved settings. The system is designed to acquire high definition images to enable visualization of capillary loops; this is a significant advantage to current, untethered ?PillCam? systems. Here, we aim to: (1) design and fabricate the ScanCap components; (2) integrate the ScanCap components and develop a tablet computer control system to display and review images; (3) evaluate performance of ScanCap in the lab and in the oral mucosa of volunteers; and (4) compare performance of ScanCap and conventional endoscopy using resected patient samples. Our multi-disciplinary team will leverage advances in consumer grade image sensors, injection molded lenses, and optical scanner technology to design a reusable capsule that rivals the performance of high-definition chromoendoscopy systems used in the US. This makes the project both high-risk, but also a game changer for the global future of esophageal endoscopy.

Abstract: Cervical cancer was once the leading cause of cancer death among women in the United States (U.S.). The implementation of comprehensive programs for screening, diagnosis, and treatment over the past 60 years has reduced cervical cancer incidence and mortality by more than 70% in the U.S. In contrast, cervical cancer remains the 1st or 2nd leading cause of cancer death among women in many low-and middle-income countries (LMICs). Cervical cancer prevention programs in low-resource settings are hampered by a lack of personnel with appropriate clinical expertise, lack of pathology services, and lack of associated infrastructure. Programs that involve multiple patient visits have a high rate of loss to follow-up. Screen-and-Treat approaches have been implemented in LMICs and include screening by HPV testing or visual inspection with acetic acid (VIA) followed by immediate treatment, reducing loss to follow-up; but these approaches lead to massive over-treatment due to the poor specificity of VIA and HPV testing. There is an urgent need for appropriate diagnostic tools to enable the implementation of a sensitive and specific Screen-Diagnose-Treat strategy that can be performed in a single patient visit in LMICs and in medically underserved areas of the U.S. We propose to develop and validate a low-cost Multimodal Mobile Colposcope (MMC) for global cervical Screen-Diagnose-Treat programs. This new device will combine the imaging capabilities of a smartphone- based colposcope developed by MobileODT with the microscopic imaging capabilities of a fiber-optic confocal imaging probe developed by Rice University. The MMC will image the entire cervix, automatically identify suspicious regions, and acquire co-registered high-resolution images of nuclear morphometry from suspicious areas. Multimodal image analysis algorithms will be developed in a study of 300 women referred for colposcopy at two sites in Brazil. The performance of the MMC with automated image analysis for detection of cervical precancer will be validated in a study of an additional 760 women referred for colposcopy in Brazil. We will determine the feasibility and usability of the MMC among low-resource setting providers by carrying out cancer prevention training courses using the MMC in Brazil. Our academic-industrial partnership includes expertise in bioengineering (Rice University), colposcopy and medical imaging (MobileODT), cervical cancer prevention and treatment (MD Anderson Cancer Center), and cervical cancer prevention and epidemiology (Albert Einstein College of Medicine), as well as the clinical expertise of our Brazilian clinical partners: Barretos Cancer Hospital and the Federal University of Health Sciences of Porto Alegre/Hospital Santa Casa. The innovative imaging technologies to be developed in this proposal will enable effective single-visit Screen-Diagnose-Treat programs to diagnose and treat women with cervical precancer while minimizing overtreatment.

Project Summary/Abstract: With over 300,000 new cases per year and a mortality rate of approximately 50%, oral cancer is a major global health issue. The stage at diagnosis is the most important predictor of survival, and unfortunately, most patients are diagnosed at a late stage. Oral cancer is preceded by visible mucosal changes which are designated oral potentially malignant disorders (OPMD). Invasive biopsy of oral lesions is the gold standard to diagnose oral dysplasia and cancer, and pathologic diagnosis of dysplasia is the best indicator of risk for oral cancer development. Dysplasia often arises in patients with OPMDs; however, most practitioners lack expertise to distinguish OPMDs from benign lesions. It is difficult even for experts to determine which oral lesions are at highest risk to contain dysplasia and should be biopsied. The goal of this proposal is to develop and validate an Active Biopsy Guidance (ABG) optical imaging system, consisting of an optical mapping scope and a high resolution microscope, to help clinicians determine precisely when and where to biopsy suspicious oral lesions. The ABG system will integrate several optical imaging modalities to non-invasively probe key molecular and morphologic changes associated with the next-generation hallmarks of cancer. In Aim 1, we will develop a compact optical mapping scope that uses Digital Light Processing technology to capture white light and auto-fluorescence images and actively project onto the oral mucosa a map highlighting areas at high risk for oral dysplasia and cancer based on loss of collagen fluorescence (a signal of invasion and metastasis) and alterations in epithelial NAD(P)H and FAD fluorescence (a signal of de-regulated cellular energetics). The mapping scope will function as the first step in the image guidance sequence, projecting a visible map of high-risk regions on the oral tissue. We will develop tracking algorithms to adjust the projected map to ensure accurate positioning despite patient movement. In Aim 2, we will develop a low-cost fluorescence and reflectance high resolution microscope capable of imaging nuclear morphology in the oral epithelium (a signal of sustained proliferative signaling, genome instability and mutation) and microvascular density and morphology (a signal of angiogenesis). The high resolution probe will serve as a confirmation modality to improve specificity. In Aim 3, we will integrate the optical mapping scope and high resolution microscope into a single, compact Active Biopsy Guidance system and validate its ability to provide real-time precise guidance for selection of oral biopsy sites in a study of high-risk patients undergoing surveillance for oral cancer. Combining widefield autofluorescence imaging and high resolution imaging of nuclear and microvessel morphology will provide a biologically directed approach to help clinicians precisely determine where and when to biopsy suspicious oral lesions, achieving both high sensitivity and high specificity. The impact of this research will be to provide interactive imaging technology that will enable earlier detection of oral neoplasia and better patient outcomes addressing a long-standing, significant global health challenge.

Abstract Oral cancer is the sixth most common malignancy worldwide. In the United States, 53,000 new cases of oral and oropharyngeal cancer are diagnosed annually. With early detection and treatment, patients with oral cancer can have excellent outcomes. However, most patients are not diagnosed until their disease is at a late stage when treatment is more invasive, more expensive, and less effective. Improving early detection of oral cancer and its precursors represents the best opportunity to reduce the incidence, morbidity, and mortality of oral cancer. In the US, rural areas are especially likely to lack effective programs for early detection of oral cancer. Early diagnosis of oral cancer in rural settings is frequently hampered by a lack of personnel with appropriate expertise, lack of health care infrastructure, limited access to health services, and long travel distances. Patients in rural counties experience longer delays and travel greater distances for diagnosis and treatment by a specialist, compared to patients in urban areas. In this project we will develop two new tools to improve early detection of oral cancer in rural areas of the US: (1) a low-cost, robust, mobile phone-based imaging system for mobile Detection of Oral Cancer (mDOC), and (2) a low-cost training model to aid in teaching oral cancer examination procedures, including the use of mDOC. These tools will provide the means to improve detection of oral cancer in rural areas through autofluorescence and white-light imaging technology, objective automated image analysis, and expert review by off-site dentists or doctors. In Aim 1 we will develop the mDOC instrument and the interactive oral exam training model. In Aim 2 we will use the mDOC device to image 120 patients referred to an oral specialist for evaluation of suspicious oral lesions. We will use this data set to develop and validate mDOC automated analysis algorithms to objectively identify high risk oral mucosal lesions. In parallel, we will conduct a usability study to evaluate and optimize the oral exam training model and the mDOC device. In Aim 3, we will use mDOC in a pilot study to image 50 patients seeking care at dental and primary care clinics in the Rio Grande Valley of south Texas, to evaluate the feasibility of oral cancer exam using mDOC in rural and underserved healthcare settings; and we will train local healthcare providers to perform oral exams using the training model and the mDOC device. Our partnership combines expertise in biomedical imaging with clinical expertise in detection, diagnosis, and treatment of oral cancer in diverse populations. The innovative mobile imaging technology to be developed in this proposal can enable scale-up of effective programs for early detection of oral cancer in rural and medically underserved regions of the United States.

Abstract Esophageal cancer is the 6th leading cause of cancer death worldwide; rates of esophageal adenocarcinoma (EAC) have risen exponentially over the past 4 decades. When EAC is diagnosed at a late stage, 5?year survival rates are dismal (?18%). Because of this, increased effort has focused on early detection and treatment of Barrett's esophagus (BE), the only recognized precursor of EAC. Indeed, when esophageal neoplasia is diagnosed at an early stage (Barrett's with high grade dysplasia/intramucosal cancer) and treated endoscopically, 5?year survival rates exceed 98%. Early detection is currently performed by standard upper endoscopy with high?definition White Light (WL) and Narrow Band Imaging (NBI) which has been shown to have a sensitivity of >90%. Nonetheless, standard endoscopy is invasive, expensive (commercial scopes >$25,000) AND requires extensive infrastructure for patient exam/sedation. Capsule endoscopy is an appealing alternative option for low?resource regions (and community practices in the US) that lack infrastructure and expert clinicians. Unfortunately, current commercially available capsule systems lack the spatial? resolution to accurately diagnose BE, are single?use, and are costly (>$25,000 per system, >$350 per single?use capsule). The goal of this proposal is to develop a lower?cost , high?resolution capsule endoscopy system to allow less experienced providers to screen for BE in community?based settings in the US and low?resource settings globally. The ScanCap system has two components: a reusable, $75 tethered capsule that is swallowed to collect images of the mucosa and $1000 supporting system to manipulate and display the collected data on a tablet computer. After the capsule is swallowed, the tether is withdrawn and high resolution, NBI images are collected from the entire esophagus. Unlike existing capsule endoscopes, our design enables high resolution side?viewing of the lumen, with circumferential scanning of the esophagus through guided rotation of the mirror within the capsule as it is withdrawn. The system is portable, battery operated, designed for use by non?physicians in underserved settings, and can acquire high definition images with NBI to enable visualization of capillary loops. Here, we aim to (1) develop and refine a tethered capsule endoscope (ScanCap) with NBI for esophageal cancer screening, (2) compare the image quality of ScanCap's NBI images to gold?standard, high?definition endoscopy with NBI using resected ex vivo samples, (3) evaluate the functional & imaging capabilities and safety of the device in an IACUC?approved porcine study, and (4) test the imaging performance of ScanCap in a pilot study of 10 patients with documented BE in an IRB?approved human study. Our multi?disciplinary team will leverage advances in consumer grade image sensors, injection molded lenses, and optical scanner technology to refine and evaluate a reusable capsule that rivals the performance of high?definition, state?of the art NBI endoscopes AND is more cost?effective (and higher?resolution) than commercial swallowed capsule systems. The end result will be a lower?cost, reusable, high quality platform for esophageal cancer screening in community?based settings in the US and rural, underserved regions worldwide.

Abstract: There is a significant need to improve global access to cervical cancer screening. Globally, approximately 570,000 women are diagnosed with and 311,000 women die from cervical cancer each year. Nearly 90 percent of cervical cancer deaths occur in low- and middle-income countries (LMICs), due mainly to challenges in implementing effective screening programs. Likewise, cervical cancer rates in medically underserved areas in the US remain high. For example, in the Rio Grande Valley of Texas, cervical cancer incidence rates are 55% higher than the US average, and approximately 10% of eligible women are screened. There is broad consensus that high-risk human papillomavirus (hrHPV) testing is the best approach to improve global screening efforts. An HPV biomarker that provides excellent sensitivity and better specificity than DNA is hrHPV mRNA. However, currently available hrHPV mRNA testing remains too complex and costly (e.g. >$45,000 for equipment and roughly $74 per test) for effective implementation into low-resource and medically underserved settings. Advances in isothermal amplification and lateral flow detection offer an opportunity to develop a point-of-care (POC) hrHPV mRNA test that is accurate, affordable, and can be performed in low-resource settings. The goal of this proposal is to combine isothermal amplification and lateral flow detection within an integrated point-of- care device to dramatically lower the cost of hrHPV mRNA testing. We will develop a low-cost, POC hrHPV E7 mRNA test that requires minimal laboratory equipment and performs as well as commercial RNA tests. Isothermal amplification reduces the instrumentation cost and complexity typically associated with nucleic acid amplification requiring only a single-temperature heater. Lateral flow detection integrates sample manipulation processes and wicks all detection reagents past pre-defined test zones, producing a simple, colorimetric readout. Our proposed proof-of-concept test will detect HPV types 16 and 18, the two types responsible for 70% of cervical cancer, and we estimate will cost <$3 per test in low-volume production. Consistent with the exploratory/developmental goals of an R21 proposal, we aim to (1) design and optimize HPV 16 and 18 E7 mRNA amplification assays and lateral flow detection; (2) combine mRNA amplification and detection into a single POC device; (3) evaluate performance of the developed mRNA test using synthetic and clinical samples. We will leverage the expertise of our interdisciplinary team, which includes designing technologies for LMICs, cervical cancer care, HPV diagnostics, and epidemiology, to develop, validate, and translate this novel screening test. We intend to build from this proof-of-concept test to incorporate HPV mRNA detection for types 31,33, 35, 45, 52, and 58 in the future to achieve detection of the HPV types that cause >90% of cervical cancer. Collectively, this research will lead to the development and implementation of a scalable, cost- effective screening test, a critical and necessary step toward the global elimination of cervical cancer.