An HER2 DNA vaccine with evolution-selected amino acid substitutions reveals a fundamental principle for cancer vaccine formulation in HER2 transgenic mice

Jones, Richard F.; Reyes, Joyce; Gibson, Heather M.; Jacob, Jennifer B.; Vaishampayan, Ulka N.; Ratner, Stuart; Chen, Kang; Wei, Wei-Zen

doi:10.1007/s00262-019-02333-9

Cited by 2 publications

(1 citation statement)

References 44 publications

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“…This cancer vaccine has been developed against the tyrosine-kinase receptor HER2, which is considered an ideal target in cancer immunotherapy due to its crucial role in the epithelial transformation and its selective overexpression in cancer tissues, such as breast cancer. − Since HER2 is exposed on the cell membrane, it can be targeted by both antibodies and cell-mediated immunity. It was previously reported that pVAX-hECTM is able to induce an antitumor immune response in breast cancer preclinical models when administered by electroporation. − Thus, we encapsulated the DNA vaccine into LNP15, while the less-effective formulation (i.e., LNP13) was used as a negative control. Immunofluorescence experiments reported in Figures and in S1 in the Supporting Information were aimed at demonstrating that transiently transfected cells can express the oncoantigen HER2 encoded by the DNA vaccine and display it on the cell surface.…”

Section: Resultsmentioning

confidence: 99%

Mechanistic Insights into the Superior DNA Delivery Efficiency of Multicomponent Lipid Nanoparticles: An In Vitro and In Vivo Study

Quagliarini

Wang

Renzi

et al. 2022

ACS Appl. Mater. Interfaces

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Lipid nanoparticles (LNPs) are currently having an increasing impact on nanomedicines as delivery agents, among others, of RNA molecules (e.g., short interfering RNA for the treatment of hereditary diseases or messenger RNA for the development of COVID-19 vaccines). Despite this, the delivery of plasmid DNA (pDNA) by LNPs in preclinical studies is still unsatisfactory, mainly due to the lack of systematic structural and functional studies on DNA-loaded LNPs. To tackle this issue, we developed, characterized, and tested a library of 16 multicomponent DNA-loaded LNPs which were prepared by microfluidics and differed in lipid composition, surface functionalization, and manufacturing factors. 8 out of 16 formulations exhibited proper size and zeta potential and passed to the validation step, that is, the simultaneous quantification of transfection efficiency and cell viability in human embryonic kidney cells (HEK-293). The most efficient formulation (LNP15) was then successfully validated both in vitro, in an immortalized adult keratinocyte cell line (HaCaT) and in an epidermoid cervical cancer cell line (CaSki), and in vivo as a nanocarrier to deliver a cancer vaccine against the benchmark target tyrosine-kinase receptor HER2 in C57BL/6 mice. Finally, by a combination of confocal microscopy, transmission electron microscopy and synchrotron small-angle X-ray scattering, we were able to show that the superior efficiency of LNP15 can be linked to its disordered nanostructure consisting of small-size unoriented layers of pDNA sandwiched between closely apposed lipid membranes that undergo massive destabilization upon interaction with cellular lipids. Our results provide new insights into the structure−activity relationship of pDNA-loaded LNPs and pave the way to the clinical translation of this gene delivery technology.

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