Many (20%-40%) cancer patients develop brain metastases, a condition with a typical life expectancy of only 3-6 months. Chemotherapy is seldom effective because systemically administered drugs reach brain metastases in very low concentrations due to (i) the blood-tumor barrier (BTB), which is created by heterogeneous vessel permeability and high interstitial pressure, (ii) the blood-brain barrier (BBB), which protects infiltrating cancer cells on tumor margins, and (iii) the nanoporous and electrostaticaly charged tissue space, which limits diffusion. Until effective strategies for overcoming these barriers emerge, brain metastasis patients will not benefit from immunotherapy. To address this problem, we have assembled a team of engineers, biologists, and clinicians with expertise in image-guided drug delivery, nanomedicine, and immunotherapy. Our goal is to engineer image-guided immunotherapies for brain metastases by combining technologies for targeted BTB/BBB opening (MRI-guided focused ultrasound and microbubbles) and improved brain-tissue penetration (nanoparticles with densely PEGylated bioinert surfaces) with strategies for enhancing T-cell infiltration and cytotoxic anti-tumor function. Our approach is supported by preliminary studies showing, for the first time, robust and sustained transgene expression in the brain via the delivery of non-viral nanoparticles with focused ultrasound. This proposal consists of 3 aims.
In Aim 1, we will develop the therapeutic delivery platform, whereby intracerebral B16 melanoma is effectively transfected via the focused ultrasound-mediated delivery of brain- penetrating nanoparticles.
Aim 1 a will be to engineer nanoparticles that effectively penetrate and transfect B16 tumors, while Aims 1b and 1c will be to develop an MRI-guided approach for safe and effective.
In Aim 2, we propose to inhibit immunosuppression and control tumor growth by the focused ultrasound-targeted delivery of signal transducer and activator of transcription 3 (STAT3)-interfering brain-penetrating nanoparticles. STAT3 is constitutively active in melanoma metastases and critical in many immunosuppressive pathways, thus STAT3 RNA interference (RNAi) should be amongst the most highly effective methods for boosting tumor immunity and inhibiting growth. Finally, in Aim 3, we propose to enhance anti-tumor immune cell infiltration and function and control tumor growth by the ultrasound-targeted delivery of brain-penetrating nanoparticles that encode an antigen recognized as foreign. In essence, our objective is to elicit an immunotherapeutic memory recall response against B16 via the high-efficiency transfection of tumor and dendritic cells with an antigen (M1 matrix protein) recognized as foreign by the host (influenza-immunized HLA-A2 mice, which recognize influenza through M1). We postulate that M1 transfection will synergize with enhanced intratumor immune cell infiltration during BBB opening to elicit a memory recall response, including the exponential expansion of potent M1-specific secondary effector T-cell populations within the tumor and deep cervical lymph nodes.

Public Health Relevance

Gene-bearing nanoparticles hold great promise for solid tumor treatment; however, targeted nanoparticle delivery is challenging. Here, we will use focused ultrasound, in conjunction with microbubbles, to target nanoparticle delivery to melanoma metastases in the brain. Nanoparticle carriers will be loaded with genes that are specifically designed to stimulate anti-tumor immune responses.

National Institute of Health (NIH)
National Cancer Institute (NCI)
Research Project (R01)
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Bioengineering, Technology and Surgical Sciences Study Section (BTSS)
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Tandon, Pushpa
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University of Virginia
Biomedical Engineering
Biomed Engr/Col Engr/Engr Sta
United States
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Mastorakos, Panagiotis; Zhang, Clark; Song, Eric et al. (2017) Biodegradable brain-penetrating DNA nanocomplexes and their use to treat malignant brain tumors. J Control Release 262:37-46
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Mead, Brian P; Kim, Namho; Miller, G Wilson et al. (2017) Novel Focused Ultrasound Gene Therapy Approach Noninvasively Restores Dopaminergic Neuron Function in a Rat Parkinson's Disease Model. Nano Lett 17:3533-3542
Zhang, Clark; Nance, Elizabeth A; Mastorakos, Panagiotis et al. (2017) Convection enhanced delivery of cisplatin-loaded brain penetrating nanoparticles cures malignant glioma in rats. J Control Release 263:112-119
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Timbie, Kelsie F; Afzal, Umara; Date, Abhijit et al. (2017) MR image-guided delivery of cisplatin-loaded brain-penetrating nanoparticles to invasive glioma with focused ultrasound. J Control Release 263:120-131
Suk, Jung Soo; Xu, Qingguo; Kim, Namho et al. (2016) PEGylation as a strategy for improving nanoparticle-based drug and gene delivery. Adv Drug Deliv Rev 99:28-51
Berry, Sneha; Mastorakos, Panagiotis; Zhang, Clark et al. (2016) Enhancing Intracranial Delivery of Clinically Relevant Non-viral Gene Vectors. RSC Adv 48:41665-41674
Mead, Brian P; Mastorakos, Panagiotis; Suk, Jung Soo et al. (2016) Targeted gene transfer to the brain via the delivery of brain-penetrating DNA nanoparticles with focused ultrasound. J Control Release 223:109-117
Mastorakos, Panagiotis; Song, Eric; Zhang, Clark et al. (2016) Biodegradable DNA Nanoparticles that Provide Widespread Gene Delivery in the Brain. Small 12:678-85

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