Tumor vasculature and microenvironment during progression and treatment : insights from optical microscopy

Thesis (Ph. D.)--Harvard-MIT Division of Health Sciences and Technology, February 2010.Vita. Cataloged from PDF version of thesis.Includes bibliographical references.In addition to cancer cells, solid tumors consist of a variety of cell types and tissues defining a complex microenvironment that influences disease progression and response to therapy. To fully characterize and probe the tumor microenvironment, new tools are needed to quantitatively assess microanatomical and physiological changes during tumor growth and treatment. Particularly important, is the metabolic microenvironment defined in tumors by hypoxia (low p02) and acidity (low pH). These parameters have been shown to influence response to radiation therapy and chemotherapy. However, very little is known about spatio-temporal changes in p02 and pH during tumor progression and therapy. By modifying the technique of intravital multiphoton microscopy (MPM) to perform phosphorescence quenching microscopy, I developed a non-invasive method to quantify oxygen tension (p02) in living tissue at high three-dimensional resolution. To probe functional changes in the metabolic microenvironment, I measured in vivo P02 during tumor growth and antiangiogenic (vascular targeted) treatment in preclinical tumor models. Nanotechnology is rapidly emerging as an important source of biocompatible tools that may shape the future of medical practice. Fluorescent semiconductor nanocrystals (NCs), also known as quantum dots, are a powerful tool for biological imaging, cellular targeting and molecular sensing.(cont.) I adapted novel fluorescence resonance energy transfer (FRET) -based nanocrystal (NC) biosensors for use with MPM to qualitatively measure in vivo extracellular pH in tumors at high-resolution. While intravital multiphoton microscopy demonstrates utility and adaptability in the study of cancer and response to therapy, the requisite high numerical aperture and exogenous contrast agents result in a limited capacity to investigate substantial tissue volumes or probe dynamic changes repeatedly over prolonged periods. By applying optical frequency domain imaging (OFDI) as an intravital microscopic tool, the technical limitations of multiphoton microscopy can be circumvented providing unprecedented access to previously unexplored, critically important aspects of tumor biology. Using entirely intrinsic mechanisms of contrast within murine tumor models, OFDI is able to simultaneously, rapidly, and repeatedly probe the microvasculature, lymphatic vessels, and tissue microstructure and composition over large volumes. Using OFDI-based techniques, measurements of tumor angiogenesis, lymphangiogenesis, tissue viability and both vascular and cellular responses to therapy were demonstrated, thereby highlighting the potential of OFDI to facilitate the exploration of pathophysiological processes and the evaluation of treatment strategies.by Ryan M. Lanning.Ph.D

Metabolic Tumor Profiling with pH, Oxygen, and Glucose Chemosensors on a Quantum Dot Scaffold

Acidity, hypoxia, and glucose levels characterize the tumor microenvironment rendering pH, pO2, and pGlucose, respectively, important indicators of tumor health. To this end, understanding how these parameters change can be a powerful tool for the development of novel and effective therapeutics. We have designed optical chemosensors that feature a quantum dot and an analyte-responsive dye. These noninvasive chemosensors permit pH, oxygen, and glucose to be monitored dynamically within the tumor microenvironment by using multiphoton imaging.National Cancer Institute (U.S.) (Grant R01-CA126642

The Personality of American Nations: An Exploratory Study

61 pagesSome scholars have presented models of the United States as a set of “nations” with distinct settlement histories and contemporary cultures. We examined personality differences in one such model, that of Colin Woodard, using data from over 75,000 respondents. Four nations were particularly distinct: The Deep South, Left Coast, New Netherland, and the Spanish Caribbean. Differences between nations at the level of the individual person were typically small, but were larger at the level of community, revealing how aggregation can contribute to differences in the lived experience of places in nations such as Yankeedom or Greater Appalachia. We represented effects in a three-dimensional model defined by Authoritarian conventionalism (which differentiated ‘Red’ and ‘Blue’ nations) as well as Cognitive resilience and Competitiveness (which differentiated among the Blue nations). Finally, we adjusted Woodard’s model to better fit the data, and found that nations largely maintained their boundaries, with the most drastic changes occurring on the East Coast

A nanocrystal-based ratiometric pH sensor for natural pH ranges

A ratiometric fluorescent pH sensor based on CdSe/CdZnS nanocrystal quantum dots (NCs) has been designed for biological pH ranges. The construct is formed from the conjugation of a pH dye (SNARF) to NCs coated with a poly(amido amine) (PAMAM) dendrimer. The sensor exhibits a well–resolved ratio response at pH values between 6 and 8 under linear or two–photon excitation, and in the presence of a 4% bovine serum albumin (BSA) solution.Chemistry and Chemical Biolog

Metabolic Tumor Profiling with pH, Oxygen, and Glucose Chemosensors on a Quantum Dot Scaffold

Acidity, hypoxia and glucose levels characterize the tumor microenvironment rendering pH, pO2 and pGlucose, respectively, important indicators of tumor health. To this end, understanding how these parameters change can be a powerful tool for the development of novel and effective therapeutics. We have designed optical chemosensors that feature a quantum dot and an analyte-responsive dye. These non-invasive chemosensors permit pH, oxygen, and glucose to be monitored dynamically within the tumor microenvironment by using multiphoton imaging.Chemistry and Chemical Biolog

The personality of American nations:An exploratory study

Some scholars have presented models of the United States as a set of “nations” with distinct settlement histories and contemporary cultures. We examined personality differences in one such model, that of Colin Woodard, using data from over 75,000 respondents. Four nations were particularly distinct: The Deep South, Left Coast, New Netherland, and the Spanish Caribbean. Differences between nations at the level of the individual person were typically small, but were larger at the level of community, revealing how aggregation can contribute to differences in the lived experience of places in nations such as Yankeedom or Greater Appalachia. We represented effects in a three-dimensional model defined by Authoritarian conventionalism (which differentiated ‘Red’ and ‘Blue’ nations) as well as Cognitive resilience and Competitiveness (which differentiated among the Blue nations). Finally, we adjusted Woodard’s model to better fit the data, and found that nations largely maintained their boundaries, with the most drastic changes occurring on the East Coast

Scaling rules for diffusive drug delivery in tumor and normal tissues

Delivery of blood-borne molecules and nanoparticles from the vasculature to cells in the tissue differs dramatically between tumor and normal tissues due to differences in their vascular architectures. Here we show that two simple measures of vascular geometry—δmax and λ—readily obtained from vascular images, capture these differences and link vascular structure to delivery in both tissue types. The longest time needed to bring materials to their destination scales with the square of δmax, the maximum distance in the tissue from the nearest blood vessel, whereas λ, a measure of the shape of the spaces between vessels, determines the rate of delivery for shorter times. Our results are useful for evaluating how new therapeutic agents that inhibit or stimulate vascular growth alter the functional efficiency of the vasculature and more broadly for analysis of diffusion in irregularly shaped domains

Metabolic Tumor Profiling with pH, Oxygen, and Glucose Chemosensors on a Quantum Dot Scaffold

Acidity, hypoxia, and glucose levels characterize the tumor microenvironment rendering pH, pO₂, and pGlucose, respectively, important indicators of tumor health. To this end, understanding how these parameters change can be a powerful tool for the development of novel and effective therapeutics. We have designed optical chemosensors that feature a quantum dot and an analyte-responsive dye. These noninvasive chemosensors permit pH, oxygen, and glucose to be monitored dynamically within the tumor microenvironment by using multiphoton imaging

The Effect of Adjuvant Trastuzumab on Locoregional Recurrence of Human Epidermal Growth Factor Receptor 2-Positive Breast Cancer Treated with Mastectomy.

BACKGROUND: Human epidermal growth factor receptor 2 (HER2) overexpression was associated with locoregional recurrence (LRR) in the preadjuvant trastuzumab era. This study aimed to examine the effect of trastuzumab on LRR in mastectomy patients and whether it varied with postmastectomy radiation (PMRT). METHODS: From the authors\u27 institutional database, 501 women with stages I-III HER2-positive breast cancer who underwent mastectomy from 1998 to 2007 were identified. A landmark analysis was performed to compare two cohorts: 170 women who received trastuzumab and 281 who did not. Kaplan-Meier methods were used to estimate locoregional recurrence-free survival (LRRFS). A propensity score analysis was used to balance the treatment groups with respect to multiple covariates. Analogous methods were used to study the effect of PMRT. RESULTS: The women in the trastuzumab group were more likely to be node positive and to receive systemic therapy or PMRT (p \u3c 0.01). The 5-year LRRFS was 98 % in the trastuzumab troup versus 94 % in the no trastuzumab group [hazard ratio (HR) 0.31; 95 % confidence interval (CI) 0.09-1.09; p = 0.07]. After adjustment for multiple covariates, including receipt of chemotherapy and PMRT, trastuzumab decreased LRR rates (HR 0.21; 95 % CI 0.04-0.94; p = 0.04). Among the women who received PMRT, trastuzumab reduced the 5-year LRR rate (0 vs 5 %; p = 0.06). Among those who did not receive PMRT, trastuzumab did not significantly decrease LRR (3 vs 6 %; p = 0.26). CONCLUSION: High rates of locoregional control (5-year rate, 98 %) were observed among patients who received trastuzumab and mastectomy ± PMRT. Trastuzumab decreased LRR in HER2-positive women who received mastectomy and PMRT, suggesting that the largest benefit is seen in a higher-risk subset of patients