Worldwide, over 2.5 million people live with spinal cord injury, with over 100,000 new cases occurring annually. Spinal cord injury often causes motor dysfunction below the level of the injury. For example, thoracic and lumbar spinal cord injury can cause paraplegia and cervical spinal cord injury can cause quadriplegia. Such injury is permanent and often severe and there is no effective treatment. Various neurologic diseases also involve damaged or dysfunctional spinal cord neurons. Neural stem cell grafts have potential for treating such conditions. However, it has not been possible to obtain sufficient numbers of appropriately patterned neural stem cells, having a spinal cord positional identity, for implanted cells to survive and functionally engraft.

The heart consists of a multitude of diverse cardiomyocyte cell types, including atrial, ventricular and pacemaker cells, which cooperate to ensure proper cardiac function and circulation throughout the body. The rhythm of the heart beat is regulated by the sinoatrial node (SAN), functionally known as the cardiac pacemaker. Loss or dysfunction of these pacemaker cardiomyocytes leads to severe cardiac arrhythmias, syncope and/or even death. Although artificial pacemakers exist to help overcome these issues, several serious limitations and problems have emerged with this approach over the past several decades including electrode fracture or damage to insulation, infection, re-operations for battery exchange, and venous thrombosis. Moreover, size mismatch and the fact that pacemaker leads do not grow with children are a concerning problem. Thus, replacing artificial pacemakers with biological pacemakers potentially overcomes these artificial pacemaker issues including the expense and complications associated with device replacement, device or lead failure, and infection. To achieve these goals, understanding how pacemaker cardiomyocytes are generated is necessary to develop a human biological pacemaker for cardiac cellular therapies.

Natural killer (NK) cells are a key component of the innate immune system and are involved in early defense against viruses and cancer cells. NK cells have the ability to lyse cells without prior sensitization  and therefore are the subject of intense interest to be potentially used as immunotherapeutic targets to treat cancer. The crucial element for using NK cells in immunotherapy is the ability to control the signaling and activation pathways. Recent work has shown that the cytokine-inducible SH2-containing protein (CIS), encoded by the Cish gene, can act as a checkpoint in NK activation by inhibiting IL-15 signaling, a major upregulator of NK cell activity. Furthermore, deletion of the Cish gene has been shown to increase the sensitivity of NK cells to IL-15, resulting in mice that are resistant to experimental metastasis.

Hematopoietic stem cell (HSC) transplants are used to treat patients with a broad spectrum of hematological malignancies, immune disorders and genetic blood diseases. Unfortunately, even after decades of use and research, there is a significant shortage of histocompatible HSCs available for transplants. Transplanting larger numbers of HSCs increases the likelihood and speed of successful engraftment, which can reduce the risk of complications such as anemia and infection, and more effectively treat underlying disease. The inability to efficiently maintain adult HSCs ex vivo is also a significant barrier for the wider development and implementation of gene therapies for diverse blood diseases and a major obstacle for engineering HSC derived cellular products for immunotherapy. One approach to overcome this challenge is to develop a means to maintain and expand HSCs in culture. Unfortunately, there is no well-defined reproducible means to maintain or expand HSCs. Even short culture times in optimized conditions are deleterious to HSCs. Ex vivo HSC maintenance and expansion could significantly enhance their clinical utility in a wide range of human diseases, providing a new platform for testing drugs, enabling more efficient gene editing within stem cells, and developing into a widely-used tool for the research community.

Rett syndrome (RTT) is a devastating disease that affects 1 in every 10,000 children born in the United States, primarily females. RTT patients undergo apparently normal development until 6-18 months of age, followed by impaired motor function, stagnation and then regression of developmental skills, hypotonia, seizures and a spectrum of autistic behaviors. Rett syndrome is a rare disease that shares certain pathways with major developmental disorders such as autism and schizophrenia, increasing the potential impact. There is no cure for Rett syndrome and the animal model does not entirely recapitulate the human disease. Thus, having the possibility to screen drugs directly in human neurons is a major milestone.

Historically, the understanding of the development and pathophysiology of the human brain has been studied by examination of post-mortem and diseased specimens in conjunction with non-human primates and mouse models. The understanding of complex biological mechanisms is driven by advancement of techniques and new model systems and recent advances in stem technologies have contributed to the advancement of our knowledge of human neural development. Moreover, the reprograming of human somatic cells into induced pluripotent stem cells (iPSCs) which can be redirected to a specific cell fate has led to a breakthrough in neurobiology research. These findings have led to the generation of human brain organoids from IPSCs.

Ocular degenerative diseases including age-related macular degeneration (AMD), retinitis pigmentosa, glaucoma, and corneal endothelial dystrophy (CED) cause irreversible vision loss and affect millions of people worldwide. Currently, there is no effective drug intervention. Grafting healthy eye cells to replenish the diseased tissues such as retina represents a promising therapeutic approach. However, previous attempts at using primary human eye cells have met with limited success due to the limited expansion capacity and differentiation potential of adult progenitors or difficulty of obtaining sufficient human fetal retinal progenitors, and possible ethical concerns. Human pluripotent stem cells (PSCs), including human embryonic stem cells (ESCs), and induced pluripotent stem cells (iPSCs) represent promising renewable donor sources for cell-based replacement therapy. Nevertheless, PSCs themselves are not suitable for direct transplantation in clinical applications due to their tendency to form teratomas and low efficiency in repopulating host tissues with desirable reprogrammed cell types in vivo. While the advancement of clinical trials of hESC-derived RPE transplants for treatment of patients with Stargardt's macular dystrophy and AMD is encouraging to the field, there is a great need for methods of generating unlimited other specialized eye cells effectively in vitro for treating blindness due to the loss of photoreceptors, RGCs and CECs. Therefore, there is a major interest in development of in vitro expandable cell sources for engineering corneal endothelium.

The researcher has developed a novel fluorescent reporter mouse in which fluorescent signals reflected endogenous Msi expression (Msi1eYFP, Msi2eGFP, Msi1 reporter mice (Reporter for Musashi1, or REM1) showed expression in the stem cell enriched adult subventricular zone, and Msi1+ cells were Nestin+ and CD133+  consistent with Msi1 marking neural stem/progenitor cells. Msi2 reporters (REM2) reflected endogenous expression of Msi2, being  highest in hematopoietic stem cells and declining with maturation.The Msi reporters described here represent exciting new tools that could be broadly useful for studying cancer. Because Msi reporter activity can be visualized through live imaging these reporter mice can be uniquely used to image and track cancer stem cells in vivo, and can provide a dynamic view of endogenous cancer growth, tumor dissemination and metastasis in its native microenvironment.  The fact that reporter positive cells are preferentially gemcitabine resistant, raises the exciting possibility that this could serve as a new platform to identify therapy resistance in vivo. The integration of such reporters in drug development may provide a powerful and sophisticated complement to traditional screens, by allowing the identification of therapies that are better able to target tumor propagating cells, and drug resistant residual disease. In addition, the spatially restricted distribution of Msi+ cells could have important implications for loco regional, aggressive targeting of driver cells that mediate resistance and disease relapse.

Cell-based therapeutics and research and development in the area of neural injury and neurological disorders could benefit from a renewable source of neural stem cells. Human embryonic stem cells provide indefinitely self-renewing cells with differentiation potential, but are inferior to lineage-restricted cells as they are prone to causing teratomas and fail to repopulate host tissues in vivo. Significant challenges in isolation and long-term cultivation of tissue-specific stem cells has restricted broad use of neural stem cells. Accordingly, there is a need for a method for obtaining a renewable source of neural stem cells.

As evidenced by fusions found in leukemia and mutations implicated in a number of myeloproliferative disorders, the JAK2 (Janus Kinase 2) gene figures prominently in blood cell development. In leukemia, it appears that JAK2 activation of Stat5A is required for tumorigenisis. In diseases with aberrant JAK2 expression or mutations, JAK2 inhibitors can be used to modulate the survival or differentiation of the affected cell population. UC researchers have explored the interaction of JAK2 and STAT5A to identify a set of markers that can be used for prognosis and monitoring of patients who may be treated with JAK2 inhibitors.

Brief description not available

Chronic Myeloid Leukemia (CML) is known to be associated with a chromosomal transposition that yields a constitutively active BCR-ABL “fusion” kinase and current therapies include kinase inhibitors (e.g., imatinib and dasatinib) that are designed to “turn off” the constitutive activation of the fusion kinase. However, these are marginally effective in the later, more aggressive stages of the disease. One cause of refractory disease is the residence of cancer stem cells (CSC) in protected tumor niches where they exit the cell cycle and revert to a quiescent state, which does not respond to the standard line of care. Such cells are found to have an altered isoform expression profile of Bcl2- family members, which may provide new means to attack stem cells that are refractory to first-line therapies.

The early (Chronic) form of Chronic Myeloid Leukemia (CML) is most commonly treated with Bcr-Abl tyrosine kinase inhibitors (e.g., imatinib and dasatinib). These drugs effectively counteract the constitutive activation of a BCR-ABL kinase, which derives from a chromosomal transposition of part of the BCR region of chromosome 22 to the ABL gene on chromosome 9. However, the Chronic phase of CML is followed by two progressively more aggressive phases and current therapies are marginally effective in the later Accelerated and Blast Crisis stages of the disease. To prevent and treat refractory forms of CML, there is a need for alternative means targeting molecular processes that fuel progression.

Despite recent advances in tissue engineering and regenerative medicine, ischemia related to cardiovascular disease results in the death of more than 100,000 amputations per year from peripheral artery disease (PAD) in the US alone.  Very few biomaterials have been examined and of those examined (e.g. fibrin, collagen, alginate, and Matrigel). None of these provide all the native components of the skeletal muscle extracellular matrix. Most are limited to improving growth factor and cell delivery. Currently no material meets all of the properties of an ideal scaffold, namely enhanced neovascularization to reduce the ischemic environment, better cell adhesion, survival, and maturation of endogenous or exogenously added cells. There is a need to develop improved compositions for minimally invasive tissue-engineered therapies for the treatment of critical limb ischemia.

Traditional chemotherapy may fail to achieve complete remission of cancers due to resistance of the underlying cancer stem cells (CSCs) to the therapeutic agents. It is now well accepted that to achieve greater efficacy there is a need to specifically target CSCs within a tumor cell population. Furthermore, understanding the self-renewal potential of these CSCs, as well as their susceptibility to drug treatment and the overall malignant potential of the cancer, are essential steps to more successful cancer therapy.

Understanding the basic mechanisms of cognitive decline and how the subcellular organization of signaling molecules is altered with cognitive decline could potentially yield novel therapeutic targets for neuronal aging and neurodegeneration.Cholesterol is a major lipid component of synapses and a limiting factor in synapse activity. Age-related impairments in the biosynthesis, transport, or uptake of cholesterol by neurons in the CNS may adversely affect synaptic circuitry. Moreover, caveolin-1 (Cav-1), a cholesterol binding and resident protein of membrane lipid rafts (MLR; discrete regions of the plasma membrane enriched in cholesterol), organizes and targets synaptic components of the neurotransmitter and neurotrophic receptor signaling pathways to MLR.

A medium formulation with the desired biological functions and that is (1) defined, (2) xeno-free with all human recombinant proteins, and (3) cost effective is crucial to the successful scale up and development of the therapeutic applications of human stem cells.

Stem cells have the potential to differentiate into a wide variety of specialized cell types. They can be used for basic research, drug discovery and, ultimately, for the treatment and prevention of disease. However, a major obstacle is that human embryonic stem (hES) cells are derived from the inner cell mass of blastocysts and derivation is encumbered by political and ethical dilemmas. Additionally, human embryonic stem cells have been found to be tumorgenic when injected into immunologically-impaired animals. Furthermore, while human embryonic stem cells potentially differentiate into multiple types of functional cells in vivo, controlled, large-scale differentiation of hES cells into specific cell types in culture has not yet been definitively demonstrated.

Brief description not available

Knowledge of DNA gene sequences and other parts of the genome of organisms has become indispensable when studying biological processes, diagnostic research, and forensic research. Following the development of dye-based sequencing methods with automated analysis, DNA sequencing has become easier and faster by a magnitude of orders. The prominent rapid high-throughput DNA sequencing methods include Genome Sequencer using pyrosequencing by Roche/454, SOliD technology by Applied Biosystems, and the sequencing by synthesis technology employed by Ilumina/Solexa. Methods for real-time direct sequencing from single DNA molecules are also emerging. These include the SMART technology being developed by Pacific Biosciences and the FRET-based sequencing method by VisiGen Biotechnologies (a part of Life Technologies Incorporated). The use of fluorescent-labeled nucleotides in almost all current single molecule-sequencing methods with optical imaging presents numerous problems, including background fluorescence and the requirement for a polymerase capable of incorporating labeled nucleotides.