Protein engineering affords researchers the unprecedented capacity to create new molecules with novel and therapeutically useful activities. Researchers have traditionally taken an unbiased approach to protein engineering, but as our knowledge of protein structure-function relationships advances, we have the exciting opportunity to apply molecular principles to guide engineering. Leveraging cutting-edge tools and exclusive expertise in structural biology and molecular design, the Spangler Lab implements a unique structure-based engineering approach to elucidate the determinants of protein activity and inform drug development.
Basic research and clinical collaborations at Johns Hopkins University empower our lab to advance novel technologies and therapeutics from the proof of concept stage all the way through the design phase, preclinical evaluation, and medical translation.
Our lab is particularly interested in engineering immune proteins, such as cytokines, antibodies, and growth factors, in order to achieve targeted activation of the immune system, either to stimulate the immune response to fight cancer or infectious diseases or to suppress the immune response for applications in autoimmune disorders, transplantation medicine, and tissue regeneration. We also build novel platforms to empower immunoengineering at the level of proteins, enabling the discovery, characterization, development, and translation of molecular technologies. Our interfacial research portfolio spans 3 key areas:
Research Area 1: Engineering cytokines and growth factors for targeted modulation of the immune response.
Due to their critical functions in regulating human physiology, cytokines and growth factors have great potential as therapeutics. However, use of natural cytokines and growth factors as drugs is limited by several factors: (i) pleiotropy, which confounds efficacy and leads to toxicity; (ii) instability, which complicates manufacturing; and (iii) short serum half-life, which limits durability. We engineer existing proteins or create entirely new proteins with enhanced selectivity, stability, and efficacy to interrogate biology and realize the potential for cytokines and growth factors as therapeutics.
We engineered a single-agent fusion protein that links interleukin-2 to an anti-cytokine antibody. The resulting immunocytokine stimulates biased expansion of regulatory T cells and protects against colitis and immune checkpoint inhibitor-induced diabetes in vivo.
We employed a hybrid computational/experimental workflow to engineer a de novo mimetic of the interleukin-4 (IL-4) cytokine that, unlike any natural protein, exclusively activates the type I IL-4 receptor. We also exploited the hyperstability of this custom-designed cytokine by directly incorporating the protein into 3D-printed biomaterial scaffolds.
Research Area 2: Designing mechanism-driven antibodies as diagnostic tools and therapeutics.
Their high affinity, specificity, advantageous pharmacological properties, and multi-layered mechanisms of action have cemented antibodies as a cornerstone of the drug development landscape. Antibodies have been engineered in a variety of formats, ranging from traditional monoclonal antibodies to multispecific antibodies and antibody fusion proteins. We design antibodies that act through various mechanisms, including steric inhibition, allosteric modulation, immune cell recruitment, and trafficking manipulation, to serve as research tools, diagnostics, and therapeutics.
We engineered a engineered a bispecific antibody that blocks signaling through the IL-6 cytokine and the IL-8 chemokine pathways to specifically and potently inhibit cancer metastasis.
We designed an antibody-enzyme fusion protein that allows for rapid, inexpensive, and precise detection of disease-associated antibody levels in human blood using commercial glucometers. We deployed this system for accurate detection of SARS-CoV-2 immunity.
Research Area 3: Building innovative molecular platforms for protein discovery and drug development.
In addition to developing cytokines, growth factors, and antibodies as research tools and potential therapeutic agents, our group is also building new platform technologies to empower the discovery, design, evaluation, and clinical advancement of proteins. The infrastructures we create at the intersection of molecular engineering, structural biophysics, and immunology advance our own research objectives, and we are also fiercely committed to sharing these resources with the greater scientific community.
We developed a framework for installing beneficial glycosylation sites into the variable domain of antibodies without disrupting function. We further demonstrated that coupling a glycoengineered antibody with a high-flux metabolite extended its in vivo half-life.
We built a suspension cell-based interaction platform (“biofloating”) that interfaces yeast-displayed antibody fragments with membrane proteins expressed on mammalian cells. We then developed a directed evolution workflow that isolated high-affinity binders against 4 distinct G protein-coupled receptors from a naive library.
© 2022 by Spangler Lab. Image credit: Matthew Blake Kelley.