James L. Elia

Over half of glioblastoma, a deadly brain cancer, do not express methylguanine methyltransferase (MGMT), an important DNA direct repair enzyme. Tumors lacking MGMT are more sensitive to specific kinds of DNA damage; however, resistance often arises by loss of mismatch repair (MMR), which senses this specific DNA damage and leads to cell death. The Bindra and Herzon Labs developed KL-50 as a novel DNA-damaging agent that is selectively toxic to MGMT-deficient cells regardless of MMR status (Lin and Gueble et al. 2022). Here, we elucidate the mechanism of the molecule in detail and offer chemical explanations for its apparent superiority over a similar DNA-damaging agent, mitozolomide (MTZ).

Shifting focus towards using the PONI polymer system for nucleic acids delivery, which already have a physiological negative charge due to the phosphate backbone. Interestingly, we found shorter PONI polymers encapsulated nucleic acids more efficiently. We used plasmid DNA delivery as a method of vaccination against Newcastle Disease Virus (NDV). We demonstrated protection against NDV within a few weeks of vaccination in a chicken model.

Continuing a nucleic acids focus from Luther et al. 2023, we used the PONI system for delivery of siRNA targeting tumor necrosis factor alpha (TNF-α), an inflammatory cytokine, to reduce damaging inflammation in the lungs. We were particularly interested in this model due to the increasing prevalence of COVID-19. Our mouse model of lung inflammation showed a strong response to the siRNA delivery at low doses, temporarily lowering TNF-α and mitigating the immune cascade.

The disadvantage of the Lee et al. 2020 E-tag system is the labor-intensive mutagenesis of each protein one wished to deliver. To address this, we made the system more modulable and accessible by biotinylating the E-tag and the protein to be delivered, then complexing both with streptavidin, a tetramer that binds tightly to biotin. Now, any protein could be delivered using our homopolymer system with “mix on the bench” chemistry, circumventing the need for extensive genetic engineering.

We wrote a tutorial review on these use of nanomaterials to enhance the efficacy and safety of transition metal catalysts (TMCs) in biological systems. We also review the potential application of nanomaterial-TMC "nanozymes" in therapeutics and imaging.

We developed a system for delivering biologics directly to the cytosol using a homopolymer called poly(oxanorbornene-imide), or “PONI,” which is made cationic via guanidinium functional groups. Our first approach used site-directed mutagenesis to add glutamates to the C-terminus of proteins (E-tag), giving the protein a bulk negative charge at physiological conditions. The negative E-tag spontaneously complexes with the positive PONI polymer in solution. With the protein encapsulated, the polymer momentarily disrupts the membrane and delivers the protein directly to the cytosol.

Presented my research at Yale Cancer Center's Radiobiology and Genome Integrity Research Seminar on the selective inhibition of homologous recombination deficient tumors using DNA-crosslinking agents.

Presented a poster on the use of a DNA-crosslinking agent with an exceptional selectivity for tumors lacking functional homologous recombination.

Preprint. See above manuscript for final version and description.

Presented my research at Yale Cancer Center's Radiobiology and Genome Integrity Research Seminar on the inhibition of ALKBH enzymes in mutant-IDH tumors by the accumulation of 2-hydroxyglutarate.

I compared the novel PONI homopolymer and E-tag system to arginine-functionalized gold nanoparticles, the previous delivery system used in the Rotello Lab. By quantifying GFP knockout in modified human cells after nanoparticle-mediated delivery of Cas9 ribonucleoprotein (RNP), we found the 60 kDa PONI homopolymer system to be superior in gene editing efficiency.

Presented my honors thesis of the same title.