Our goal is to develop life‐saving tools for the critically‐injured patient. Over the years, we havetranslated a technique developed at UC Irvine, Diffuse Optical Spectroscopic Imaging (DOSI), from theresearch bench to the cusp of a 5‐center clinical trial designed to monitor the effects of neoadjuvantchemotherapy. These same technologies and approaches that have been effective in measuring breasttumor metabolic response to therapy could also be used to detect early hemorrhage well beforetraditional clinical markers (such as arterial blood pressure) give notice. This could be accomplished bysensing the metabolic/hemodynamic status of mucosal tissues, such as in the esophagus.
However, there are challenges moving DOSI from external to internal tissues, such as developing asimple‐use DOSI probe which is compatible with minimally invasive medical tools. We haveconceptualized a method that integrates standard critical‐care tools (nasogastric tube) with DOSI opticalfibers to quantify mucosal tissue oxygenation and blood volume. This probe must be constructed andtested, and would provide exciting preliminary data for both clinical and technological proposals. Oncethe barriers for DOSI to minimally invasive tools has been overcome, new medical applications (such asenhanced GI cancer detection), can be pursued.
Melanoma is an aggressive tumor resistant to both chemotherapy and immunotherapy1. While molecularlytargeted therapies have shown promise in shrinking select melanomas, a proportion of these tumors stillrecur2. We recently utilized a genome-wide synthetic lethal RNAi screening approach to uncover the molecularfulcrums that allow melanoma cells to be resistant to conventional and molecularly-directed therapies. Ourscreen identified three less studied Rho GTPases as central regulators of melanoma chemoresistance.Functional assays revealed that one of these GTPases, RhoJ, controls melanoma migration, invasion, andapoptotic resistance. Followup studies revealed that RhoJ controls the expression of Sox10, a neural cresttranscription factor known to regulate melanoblast migration and stem cell maintenance in the developingembryo3. In this proposal, we seek to translate our screen results from the bench to the bedside by directlymeasuring the expression of Sox10 in melanoma tumors from patients currently on melanoma clinical trials atUC Irvine. As a result of this funding, we will create translational resources for future studies on melanomachemoresistance and generate critical preliminary data for an R01 application to be submitted in March thatfocuses on the role of RhoJ and Sox10 in melanoma chemoresistance.
Facioscapulohumeral (FSH) muscular dystrophy is a common autosomal dominant neuromuscular disease for which there is no effective treatment. A very recent study suggests that skeletal muscle regeneration in FSH is disrupted by a gain-of-function mutation that produces a toxic gene product. The mutation creates a poly A signal that stabilizes transcripts of DUX4, a double homeobox gene of unknown function. We propose that correction of this mutation could revert FSH cells to wildtype. Previously, we developed induced pluripotent stem cells (iPS) cells from FSH patients. The new finding about DUX4 expression in FSH provides a unique opportunity to correct the mutation in FSH cells as a route to clinical application, i.e. cell-based therapy. We propose to normalize the mutant allele in FSH-iPS cells using a powerful new approach. The technology uses zinc finger nucleases (ZFNs) to make a defined genetic change at a specified site in the genome. Because iPS cells can give rise to differentiated cells, e.g. muscle, iPS cells from FSH patients could be corrected and used to replace damaged muscle in FSH patients. Demonstration of our ability to correct the mutation in FSH patients would allow us to apply for upcoming NIH and CIRM funding for translational research.
The goal of this project is to accelerate the translation of a laboratory finding into clinicalapplications using choroid plexus epithelial cells (CPECs). CPECs nourish, maintain,and protect the central nervous system (CNS) via cerebrospinal fluid (CSF) secretionand blood-CSF barrier formation. Moreover, CPEC dysfunction contributes to manyneurodegenerative disorders, with some even postulating a causal role in Alzheimer’sdisease, and choroid plexus transplants have been effective in animal models ofHuntington’s disease and stroke. These and other normal roles, disease associations,and transplant studies provide significant rationale for CPEC engineering, replacement,and transplant applications, but development of these applications have been stymied bythe inability to generate or expand CPECs in culture. We believe that this roadblock hasnow been overcome, based on our evidence for CPEC generation from mouse andhuman embryonic stem cells (ESCs) (manuscript submitted, provisional patentapplication pending). The specific goal of this ICTS proposal is to develop afluorescence-activated cell sorting (FACS) method for CPEC enrichment from humanESC cultures. If successful, this will accelerate the translation of our findings into clinicalapplications using CPECs and a brand-new area of regenerative medicine.
Insulin resistance is a common problem for which there are no safe and effective pharmacological treatments. Insulin resistance is associated with heart disease, stroke, cancer and non-alcoholic fatty liver disease (NAFLD), a condition that afflicts more than 40% of US adults and can lead to cirrhosis and liver cancer. Betaine, a normal cellular component, is sold OTC as a nutritional supplement. Betaine has been studied in humans with elevated serum homocysteine and reported to be safe. We found that oral betaine eliminated insulin resistance and reduced liver fat when fed to mice with NAFLD. Given the effectiveness in animal models and safety in humans, we propose to test betaine as a treatment for insulin resistance in 15 subjects with fasting blood sugar between 100 and 125 mg/dL. Subjects will be given placebo, betaine 4g/day, 8g/day and 12g/day, for one month at each dose. Our primary outcome is change in homeostasis model assessment for insulin resistance (HOMA-IR) score at the end of treatment as compared with the beginning. We will also measure changes in serum lipids as well as markers of oxidative stress and inflammation. This study has been approved by the IRB and exempted by the FDA.
Spinal muscular atrophy (SMA), an autosomal recessive disorder of lower motor neurons, is classifiedby age of onset and walking ability. Two almost identical survival motor neuron (SMN) genes exist onchromosome 5q13. SMN1 forms full length transcript and homozygous deletions of exons 7 and 8 inSMN1 are present in 95% of SMA patients. SMN2 forms truncated transcripts due to exon 7skipping. Previous work indicates that the SMN2 copy number dictates disease onset andseverity of progression. We propose to extend these findings by examining the phenotypic contributionSMN2 alternative pre-mRNA splicing patterns. SMA with calf hypertrophy is an uncommon, but milderSMA phenotype with no current information about the role of SMN 2 copy number or disease modifiers.There is also limited imaging data available for these patients. We have determined that we have thepatient population that merits a pilot study to evaluate SMN2 copy number, alternative splicingpatterns and MRI in SMA patients with calf hypertrophy with the goal to identify predictors ofphenotype and biomarkers for natural history studies and future clinical trials. Our findings may lead topreliminary data for external funding and may identify novel targets for drug therapy.
This proposal seeks funds for a pilot study to establish the utility of multiplexing with quantum dots labeling biomarkers on Formalin-Fixed Paraffin-Embedded Tissues for prediction of prostate cancer recurrence and metastasis after prostatectomy. In addition, we will evaluate the pharmacokinetics of a first-in-class humanized wnt inhibitory factor-1 (WIF1) recombinant protein in mice to establish the doses and routes (i.e.: i.v. vs. i.p. injection) for WIF1-human IgG and anti-VEGF antibody combined therapy for advanced prostate cancer in preclinical studies. With completion of these two pilot studies, we plan to submit an R01 application to test our novel hypothesis: anti-VEGF therapies, while blocking VEGF and VEGF receptors, produce hypoxic conditions and activate the Wnt pathway leading to epithelial-to-mesenchymal transition (EMT) and increasing the expression of HGF/c-Met and co-receptor Nuropilin-2 (NRP2) and, therefore, the interaction between NRP2 and c-Met. Collectively, these molecular events by hypoxia cause cancer cell migration and invasion, eventually resulting tumor metastasis and disease progression. Our recently published and unpublished data have provided evidence for a therapeutic approach that WIF1 inhibits Wnt signaling leading to a reversal of EMT and down-regulation of NRP2 and c-Met, which prevent anti-VEGF therapies-induced cancer metastasis and progression.