The objective of this project is to create a microfluidic platform capable of characterizing the cellular and cytokine content of vitreous humor. This platform will be of direct benefit to patients suffering from uveitis or primary intraocular lymphoma (PIOL) who need to undergo surgical removal of the vitreous humor (vitreous biopsy) in order to obtain a detailed diagnosis of these diseases. Microfluidic immunophenotyping of vitreous humor is an attractive alternative to the current approaches of cytology and flow cytometry because it requires considerably fewer cells and allows rapid analysis of cells that are typically very fragile.
Both uveitis and PIOL are diseases associated with the invasion of lymphocytes into various regions of the eye. Approximately 2.3 million Americans suffer from uveitis, and vision-loss due to uveitis accounts for 10-15% of blindness in the U.S. PIOL is manifested by the presence of malignant B-lymphocytes which may indicate tumor formation in the brain, retina, or other areas of the central nervous system. Although PIOL is an uncommon disease, patients of PIOL have a low likelihood of recovery because of the lack of early detection techniques.
The vitreous biopsy is performed to distinguish PIOL from uveitis and identify a targeted therapeutic path. For PIOL, early detection allows patients to begin chemo- and radio-therapy. For uveitis, identification of its underlying cause allows treatement with specific drugs instead of broad-target immunosuppressive agents that typically cause severe side effects. The current approach to analyze the vitreous biopsy product provides a conclusive diagnosis in only 15% of patients because it is constrained by the small number of cells available for analysis and their fragility.
The following tasks will be carried out to accomplish the project objective:
- Design and fabricate microfluidic devices for cell and cytokine capture based on antibody-antigen interactions. - Create a microfluidic immunophenotyping technique based on the chemotactic deflection of cells using gradients of dissolved antibodies. - Characterize the performance of the microfluidic devices using a model that mimics the cellular and cytokine content of vitreous humor extracted from patients of uveitis or PIOL. - Determine lower bounds of target cell and cytokine concentration for the multiple markers of each disease type. - Immunophenotype cells and cytokines in vitreous fluid obtained from patient vitreous biopsy samples.
Intellectual Merit: If successful, the proposed research will provide a new and transformative method to rapidly identify cells and cytokines associated with uveitis and PIOL, overcoming the shortcomings of current analysis techniques. Part of the proposed microfluidic approach is technologically transformative gradient-based design to identify multiple cell surface markers on individual cells without requiring any labeling. The ability to determine cell- and cytokine-type from very small sample sizes will allow for early diagnosis of PIOL, which cannot be performed using current techniques. The PI has worked in the area of microfluidic cell analysis for over 5 years and this project is a collaborative effort between with Dr. Kameran Laskhari, an ophthalmic surgeon at the Schepens Eye Research Institute in Boston.
Broader Impact: (a) If successful, the proposed research will provide an accurate and rapid diagnosis technique for patients with uveitis or PIOL. (b) The proposed research will involve high school students and science teachers in experimental research via existing Young Scholar and RET programs at Northeastern University, and (c) involve undergraduates in experimental research. (d) High school students participating in the proposed research will be encouraged to develop science fair projects and pursue careers in biomedical science and engineering. (e) Disseminate research results and describe implications for uveitis and PIOL therapeutics through a website.
The objective of this project was to create devices capable of detecting the protein and cell content of biopsy samples taken from the eye with the focus on two diseases, uveitis and intraocular lymphoma. Such analysis of fluid samples from the eye is difficult because the concentrations of protein molecules and cells in these samples are extremely low. Furthermore, there is a general need to make such analysis easy to perform at doctor's offices to reduce the cost and time associated with off-site labs, and allow physicians to make treatment decisions more quickly. This research addresses the need for innovative diagnostic techniques to aid ophthalmologists in distinguishing between ocular cancer and uveitis. Ocular biopsies are unusually large in size and therefore difficult to characterize using current technology. The goal of this research is to create a microfluidic platform capable of providing a comprehensive analysis of ocular biopsies. A detailed diagnosis is critical for the early detection of ocular cancer as well as for identifying targeted therapeutic drugs to treat uveitis. The research: * has resulted in a microfluidic device capable of characterizing the cytokine content of the eye (see accompanying image). The device will be implemented in the clinical setting in order to map the full cytokine profiles of uveitis and ocular cancer patients to discover possible correlations between cytokine presence and disease type. * has initiated the integration of microfluidics into ocular medical diagnostic devices for point-of-care application. While considerably more work must be done to validate and make this technology deployable in clinics and economically viable, support from this National Science Foundation award was nevertheless instrumental in enabling the initial foundational work for such systems.
