Bio

Honors & Awards


  • Stanford Graduate Fellowship, Stanford University (2018-2021)

Education & Certifications


  • BS, University of Washington, Biochemistry (2015)

Publications

All Publications


  • Engineering Protein-Secreting Plasma Cells by Homology-Directed Repair in Primary Human B Cells MOLECULAR THERAPY Hung, K. L., Meitlis, I., Hale, M., Chen, C., Singh, S., Jackson, S. W., Miao, C. H., Khan, I. F., Rawlings, D. J., James, R. G. 2018; 26 (2): 456–67

    Abstract

    The ability to engineer primary human B cells to differentiate into long-lived plasma cells and secrete a de novo protein may allow the creation of novel plasma cell therapies for protein deficiency diseases and other clinical applications. We initially developed methods for efficient genome editing of primary B cells isolated from peripheral blood. By delivering CRISPR/CRISPR-associated protein 9 (Cas9) ribonucleoprotein (RNP) complexes under conditions of rapid B cell expansion, we achieved site-specific gene disruption at multiple loci in primary human B cells (with editing rates of up to 94%). We used this method to alter ex vivo plasma cell differentiation by disrupting developmental regulatory genes. Next, we co-delivered RNPs with either a single-stranded DNA oligonucleotide or adeno-associated viruses containing homologous repair templates. Using either delivery method, we achieved targeted sequence integration at high efficiency (up to 40%) via homology-directed repair. This method enabled us to engineer plasma cells to secrete factor IX (FIX) or B cell activating factor (BAFF) at high levels. Finally, we show that introduction of BAFF into plasma cells promotes their engraftment into immunodeficient mice. Our results highlight the utility of genome editing in studying human B cell biology and demonstrate a novel strategy for modifying human plasma cells to secrete therapeutic proteins.

    View details for DOI 10.1016/j.ymthe.2017.11.012

    View details for Web of Science ID 000425206300014

    View details for PubMedID 29273498

    View details for PubMedCentralID PMC5835153

  • Wnt signaling and tbx16 form a bistable switch to commit bipotential progenitors to mesoderm DEVELOPMENT Bouldin, C. M., Manning, A. J., Peng, Y., Farr, G. H., Hung, K. L., Dong, A., Kimelman, D. 2015; 142 (14): 2499-+

    Abstract

    Anterior to posterior growth of the vertebrate body is fueled by a posteriorly located population of bipotential neuro-mesodermal progenitor cells. These progenitors have a limited rate of proliferation and their maintenance is crucial for completion of the anterior-posterior axis. How they leave the progenitor state and commit to differentiation is largely unknown, in part because widespread modulation of factors essential for this process causes organism-wide effects. Using a novel assay, we show that zebrafish Tbx16 (Spadetail) is capable of advancing mesodermal differentiation cell-autonomously. Tbx16 locks cells into the mesodermal state by not only activating downstream mesodermal genes, but also by repressing bipotential progenitor genes, in part through a direct repression of sox2. We demonstrate that tbx16 is activated as cells move from an intermediate Wnt environment to a high Wnt environment, and show that Wnt signaling activates the tbx16 promoter. Importantly, high-level Wnt signaling is able to accelerate mesodermal differentiation cell-autonomously, just as we observe with Tbx16. Finally, because our assay for mesodermal commitment is quantitative we are able to show that the acceleration of mesodermal differentiation is surprisingly incomplete, implicating a potential separation of cell movement and differentiation during this process. Together, our data suggest a model in which high levels of Wnt signaling induce a transition to mesoderm by directly activating tbx16, which in turn acts to irreversibly flip a bistable switch, leading to maintenance of the mesodermal fate and repression of the bipotential progenitor state, even as cells leave the initial high-Wnt environment.

    View details for DOI 10.1242/dev.124024

    View details for Web of Science ID 000358294200013

    View details for PubMedID 26062939

    View details for PubMedCentralID PMC4510867