TMC Biodesign: The Design and Implementation of a Product Development Framework for Successful Innovation in the Healthcare Industry.

It is not uncommon to see both academic and industry institutions speed through, or even outright skip, the different stages of innovation. Industry often considers early stages of innovation, such as needs identification, to be too risky, or a waste of time and resources. They tend to focus more on improving validated solutions and creating incremental changes, resulting in products that lack innovation. Academia often considers aspects of the innovation process to be too commercial to consider during their research initiatives, which often results in the development of great technologies that cannot be implemented due to their lack of commercial viability, resulting in a great deal of wasted time and capital. There is a stark need to train everyone involved in the product development process to properly appreciate and implement all stages of the innovation cycle. Engineers, physicians, and business-minded people need to be taught how to come together to solve healthcare’s biggest problems. They need to learn how to turn technological developments into commercially viable products that solve customer needs. In partnership with the Texas Medical center, I present in this research a framework for providing future medical technology leaders the experience required to create transformational solutions to healthcare’s biggest challenges. I provide a structured process for innovating in the complex healthcare industry, beginning with first-hand observations of clinical needs and ending with a plan for commercializing a medical product. This thesis is intended to describe the proposed framework for medical device innovation and evaluate its potential for success through participation in the inaugural fellowship

TMC Biodesign: The Design and Implementation of a Product Development Framework for Successful Innovation in the Healthcare Industry.

It is not uncommon to see both academic and industry institutions speed through, or even outright skip, the different stages of innovation. Industry often considers early stages of innovation, such as needs identification, to be too risky, or a waste of time and resources. They tend to focus more on improving validated solutions and creating incremental changes, resulting in products that lack innovation. Academia often considers aspects of the innovation process to be too commercial to consider during their research initiatives, which often results in the development of great technologies that cannot be implemented due to their lack of commercial viability, resulting in a great deal of wasted time and capital. There is a stark need to train everyone involved in the product development process to properly appreciate and implement all stages of the innovation cycle. Engineers, physicians, and business-minded people need to be taught how to come together to solve healthcare’s biggest problems. They need to learn how to turn technological developments into commercially viable products that solve customer needs. In partnership with the Texas Medical center, I present in this research a framework for providing future medical technology leaders the experience required to create transformational solutions to healthcare’s biggest challenges. I provide a structured process for innovating in the complex healthcare industry, beginning with first-hand observations of clinical needs and ending with a plan for commercializing a medical product. This thesis is intended to describe the proposed framework for medical device innovation and evaluate its potential for success through participation in the inaugural fellowship

Single-Cell Transcriptional Analysis of Normal, Aberrant, and Malignant Hematopoiesis in Zebrafish

Hematopoiesis culminates in the production of functionally heterogeneous blood cell types. In zebrafish, the lack of cell surface antibodies has compelled researchers to use fluorescent transgenic reporter lines to label specific blood cell fractions. However, these approaches are limited by the availability of transgenic lines and fluorescent protein combinations that can be distinguished. Here, we have transcriptionally profiled single hematopoietic cells from zebrafish to define erythroid, myeloid, B, and T cell lineages. We also used our approach to identify hematopoietic stem and progenitor cells and a novel NK-lysin 4+ cell type, representing a putative cytotoxic T/NK cell. Our platform also quantified hematopoietic defects in rag2E450fs mutant fish and showed that these fish have reduced T cells with a subsequent expansion of NK-lysin 4+ cells and myeloid cells. These data suggest compensatory regulation of the innate immune system in rag2E450fs mutant zebrafish. Finally, analysis of Myc-induced T cell acute lymphoblastic leukemia showed that cells are arrested at the CD4+/CD8+ cortical thymocyte stage and that a subset of leukemia cells inappropriately reexpress stem cell genes, including bmi1 and cmyb. In total, our experiments provide new tools and biological insights into single-cell heterogeneity found in zebrafish blood and leukemia

Notch signaling enhances bone regeneration in the zebrafish mandible

Loss or damage to the mandible caused by trauma, treatment of oral malignancies, and other diseases is treated using bone-grafting techniques that suffer from numerous shortcomings and contraindications. Zebrafish naturally heal large injuries to mandibular bone, offering an opportunity to understand how to boost intrinsic healing potential. Using a novel her6:mCherry Notch reporter, we show that canonical Notch signaling is induced during the initial stages of cartilage callus formation in both mesenchymal cells and chondrocytes following surgical mandibulectomy. We also show that modulation of Notch signaling during the initial post-operative period results in lasting changes to regenerate bone quantity one month later. Pharmacological inhibition of Notch signaling reduces the size of the cartilage callus and delays its conversion into bone, resulting in non-union. Conversely, conditional transgenic activation of Notch signaling accelerates conversion of the cartilage callus into bone, improving bone healing. Given the conserved functions of this pathway in bone repair across vertebrates, we propose that targeted activation of Notch signaling during the early phases of bone healing in mammals may both augment the size of the initial callus and boost its ossification into reparative bone.</p

High-throughput cell transplantation establishes that tumor-initiating cells are abundant in zebrafish T-cell acute lymphoblastic leukemia

Self-renewal is a feature of cancer and can be assessed by cell transplantation into immune-compromised or immune-matched animals. However, studies in zebrafish have been severely limited by lack of these reagents. Here, Myc-induced T-cell acute lymphoblastic leukemias (T-ALLs) have been made in syngeneic, clonal zebrafish and can be transplanted into sibling animals without the need for immune suppression. These studies show that self-renewing cells are abundant in T-ALL and comprise 0.1% to 15.9% of the T-ALL mass. Large-scale single-cell transplantation experiments established that T-ALLs can be initiated from a single cell and that leukemias exhibit wide differences in tumor-initiating potential. T-ALLs also can be introduced into clonal-outcrossed animals, and T-ALLs arising in mixed genetic backgrounds can be transplanted into clonal recipients without the need for major histocompatibility complex matching. Finally, high-throughput imaging methods are described that allow large numbers of fluorescent transgenic animals to be imaged simultaneously, facilitating the rapid screening of engrafted animals. Our experiments highlight the large numbers of zebrafish that can be experimentally assessed by cell transplantation and establish new high-throughput methods to functionally interrogate gene pathways involved in cancer self-renewal

Clonal fate mapping quantifies the number of haematopoietic stem cells that arise during development

Haematopoietic stem cells (HSCs) arise in the developing aorta during embryogenesis. The number of HSC clones born has been estimated through transplantation, but experimental approaches to assess the absolute number of forming HSCs in a native setting have remained challenging. Here, we applied single-cell and clonal analysis of HSCs in zebrafish to quantify developing HSCs. Targeting creERT2 in developing cd41:eGFP+ HSCs enabled long-term assessment of their blood contribution. We also applied the Brainbow-based multicolour Zebrabow system with drl:creERT2 that is active in early haematopoiesis to induce heritable colour barcoding unique to each HSC and its progeny. Our findings reveal that approximately 21 HSC clones exist prior to HSC emergence and 30 clones are present during peak production from aortic endothelium. Our methods further reveal that stress haematopoiesis, including sublethal irradiation and transplantation, reduces clonal diversity. Our findings provide quantitative insights into the early clonal events that regulate haematopoietic development