Self‐healing encapsulation and controlled release of vaccine antigens from PLGA microparticles delivered by microneedle patches

There is an urgent need to reduce reliance on hypodermic injections for many vaccines to increase vaccination safety and coverage. Alternative approaches include controlled release formulations, which reduce dosing frequencies, and utilizing alternative delivery devices such as microneedle patches (MNPs). This work explores development of controlled release microparticles made of poly (lactic‐co‐glycolic acid) (PLGA) that stably encapsulate various antigens though aqueous active self‐healing encapsulation (ASE). These microparticles are incorporated into rapid‐dissolving MNPs for intradermal vaccination.PLGA microparticles containing Alhydrogel are loaded with antigens separate from microparticle fabrication using ASE. This avoids antigen expsoure to many stressors. The microparticles demonstrate bi‐phasic release, with initial burst of soluble antigen, followed by delayed release of Alhydrogel‐complexed antigen over approximately 2 months in vitro. For delivery, the microparticles are incorporated into MNPs designed with pedestals to extend functional microneedle length. These microneedles readily penetrate skin and rapidly dissolve to deposit microparticles intradermally. Microparticles remain in the tissue for extended residence, with MNP‐induced micropores resealing readily. In animal models, these patches generate robust immune responses that are comparable to conventional administration techniques. This lays the framework for a versatile vaccine delivery system that could be self‐applied with important logistical advantages over hypodermic injections.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/147859/1/btm210103-sup-0001-supinfo.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/147859/2/btm210103_am.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/147859/3/btm210103.pd

Immunotherapy for gliomas: shedding light on progress in preclinical and clinical development

Gliomas are infiltrating brain tumors associated with high morbidity and mortality. Current standard of care includes radiation, chemotherapy and surgical resection. Today, survival rates for malignant glioma patients remain dismal and unchanged for decades. The glioma microenvironment is highly immunosuppressive and consequently this has motivated the development of immunotherapies for counteracting this condition, enabling the immune cells within the tumor microenvironment to react against this tumor.Areas covered: The authors discuss immunotherapeutic strategies for glioma in phase-I/II clinical trials and illuminate their mechanisms of action, limitations and key challenges. They also examine promising approaches under preclinical development.Expert opinion: In the last decade there has been an expansion in immune-mediated anti-cancer therapies. In the glioma field, sophisticated strategies have been successfully implemented in preclinical models. Unfortunately, clinical trials have not yet yielded consistent results for glioma patients. This could be attributed to our limited understanding of the complex immune cell infiltration and its interaction with the tumor cells, the selected time for treatment, the combination with other therapies and the route of administration of the agent. Applying these modalities to treat malignant glioma is challenging, but many new alternatives are emerging to by-pass these hurdles.Fil: Garcia Fabiani, Maria Belen. University of Michigan; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Ventosa, Maria. University of Michigan; Estados UnidosFil: Comba, Andrea. University of Michigan; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Candolfi, Marianela. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas; ArgentinaFil: Nicola Candia, Alejandro Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas; ArgentinaFil: Alghamri, Mahmoud S.. University of Michigan; Estados UnidosFil: Kadiyala, Padma. University of Michigan; Estados UnidosFil: Carney, Stephen. University of Michigan; Estados UnidosFil: Faisal, Syed M.. University of Michigan; Estados UnidosFil: Schwendeman, Anna. University of Michigan; Estados UnidosFil: Moon, James J.. University of Michigan; Estados UnidosFil: Scheetz, Lindsay. University of Michigan; Estados UnidosFil: Lahann, Joerg. University of Michigan; Estados UnidosFil: Mauser, Ava. University of Michigan; Estados UnidosFil: Lowenstein, Pedro R.. University of Michigan; Estados UnidosFil: Castro, Maria Gabriela. University of Michigan; Estados Unido

Genetic Alterations in Gliomas Remodel the Tumor Immune Microenvironment and Impact Immune-Mediated Therapies

High grade gliomas are malignant brain tumors that arise in the central nervous system, in patients of all ages. Currently, the standard of care, entailing surgery and chemo radiation, exhibits a survival rate of 14-17 months. Thus, there is an urgent need to develop new therapeutic strategies for these malignant brain tumors. Currently, immunotherapies represent an appealing approach to treat malignant gliomas, as the pre-clinical data has been encouraging. However, the translation of the discoveries from the bench to the bedside has not been as successful as with other types of cancer, and no long-lasting clinical benefits have been observed for glioma patients treated with immune-mediated therapies so far. This review aims to discuss our current knowledge about gliomas, their molecular particularities and the impact on the tumor immune microenvironment. Also, we discuss several murine models used to study these therapies pre-clinically and how the model selection can impact the outcomes of the approaches to be tested. Finally, we present different immunotherapy strategies being employed in clinical trials for glioma and the newest developments intended to harness the immune system against these incurable brain tumors.Fil: Garcia Fabiani, Maria Belen. University of Michigan; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Haase, Santiago. University of Michigan; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Comba, Andrea. University of Michigan; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Carney, Stephen. University of Michigan; Estados UnidosFil: McClellan, Brandon. University of Michigan; Estados UnidosFil: Banerjee, Kaushik. University of Michigan; Estados UnidosFil: Alghamri, Mahmoud S.. University of Michigan; Estados UnidosFil: Syed, Faisal. University of Michigan; Estados UnidosFil: Kadiyala, Padma. University of Michigan; Estados UnidosFil: Nuñez, Felipe. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaFil: Candolfi, Marianela. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas; ArgentinaFil: Asad, Antonela Sofía. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas; ArgentinaFil: González, Nazareno. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Biomédicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Biomédicas; ArgentinaFil: Aikins, Marisa E.. University of Michigan; Estados UnidosFil: Schwendeman, Anna. University of Michigan; Estados UnidosFil: Moon, James J.. University of Michigan; Estados UnidosFil: Lowenstein, Pedro R.. University of Michigan; Estados UnidosFil: Castro, Maria G.. University of Michigan; Estados Unido

31st Annual Meeting and Associated Programs of the Society for Immunotherapy of Cancer (SITC 2016) : part two

Background The immunological escape of tumors represents one of the main ob- stacles to the treatment of malignancies. The blockade of PD-1 or CTLA-4 receptors represented a milestone in the history of immunotherapy. However, immune checkpoint inhibitors seem to be effective in specific cohorts of patients. It has been proposed that their efficacy relies on the presence of an immunological response. Thus, we hypothesized that disruption of the PD-L1/PD-1 axis would synergize with our oncolytic vaccine platform PeptiCRAd. Methods We used murine B16OVA in vivo tumor models and flow cytometry analysis to investigate the immunological background. Results First, we found that high-burden B16OVA tumors were refractory to combination immunotherapy. However, with a more aggressive schedule, tumors with a lower burden were more susceptible to the combination of PeptiCRAd and PD-L1 blockade. The therapy signifi- cantly increased the median survival of mice (Fig. 7). Interestingly, the reduced growth of contralaterally injected B16F10 cells sug- gested the presence of a long lasting immunological memory also against non-targeted antigens. Concerning the functional state of tumor infiltrating lymphocytes (TILs), we found that all the immune therapies would enhance the percentage of activated (PD-1pos TIM- 3neg) T lymphocytes and reduce the amount of exhausted (PD-1pos TIM-3pos) cells compared to placebo. As expected, we found that PeptiCRAd monotherapy could increase the number of antigen spe- cific CD8+ T cells compared to other treatments. However, only the combination with PD-L1 blockade could significantly increase the ra- tio between activated and exhausted pentamer positive cells (p= 0.0058), suggesting that by disrupting the PD-1/PD-L1 axis we could decrease the amount of dysfunctional antigen specific T cells. We ob- served that the anatomical location deeply influenced the state of CD4+ and CD8+ T lymphocytes. In fact, TIM-3 expression was in- creased by 2 fold on TILs compared to splenic and lymphoid T cells. In the CD8+ compartment, the expression of PD-1 on the surface seemed to be restricted to the tumor micro-environment, while CD4 + T cells had a high expression of PD-1 also in lymphoid organs. Interestingly, we found that the levels of PD-1 were significantly higher on CD8+ T cells than on CD4+ T cells into the tumor micro- environment (p < 0.0001). Conclusions In conclusion, we demonstrated that the efficacy of immune check- point inhibitors might be strongly enhanced by their combination with cancer vaccines. PeptiCRAd was able to increase the number of antigen-specific T cells and PD-L1 blockade prevented their exhaus- tion, resulting in long-lasting immunological memory and increased median survival

MICROWAVE SPECTRA AND STRUCTURES OF AMINODIFLUORO PHOSPHINE AND CHLORODIFLUOROPHOSPHINE

Author Institution: Department of Chemistry, Michigan State UniversityThe microwave spectra of $PF_{2}^{14} NH_{1}$, $PF_{2}^{12} NH_{2}$, $PF_{2}NHD$^{\prime} and $PF_{2}NDH$^{\prime} have been assigned and from these the structure of the molecule has been determined. The phosphorous, nitrogen and hydrogen atoms appear to lie in the symmetry plane of the molecule and the structural parameters obtained are: $r_{PF}=1.586$ {\AA}, $r_{PN}=1.647$ {\AA} $r_{NH(cis)}=1.000$ {\AA}, $r_{NH(trass)}=0.983$ {\AA}, $<{FPF}=94.7^\circ$. $<{FPN}=100.7^\circ$, $<{HNH}=117.6^\circ$, $< PNH_{cis}^\circ=122.9^\circ$ and $< PNH_{tanss}=119.5^\circ$. The dipole moment components are found to be $\mu_{s}=2.57 D$ and $\mu_{e}=0.18 D$ giving a total dipole $\mu$=2.58D. Analysis of the hyperfine structure on transitions for $PF_{2}^{14} NH_{2}$ yielded the quadrupole coupling constants $\chi_{aa} = 1.65$ MHz, $\chi_{bb} = -3.45$ MHz and $\chi_{cc} = 1.80$ MHz. Torsional satellites have been seen and the results of an investigation of them will be reported. The structure of chlorodifluorophosphine was determined following the assignment of the microwave spectra of $PF_{2}^{23}Cl$ and $PF_{2}^{27}Cl$. The structural parameters are: $r_{PF}=1.571$ \AA, $r_{PCI}=2.030$ \AA. $<{FPF}=97.3^\circ$ and $<{FPCl}=99.2^\circ$. The dipole moment components are determined to be $\mu_{a}=0.11 D$ and $\mu_{e}$=0.87D giving a total dipole $\mu$=0.88D. The quadrupole coupling constants in $PF_{2}^{25}Cl$ are $\chi_{aa}= -39.3$ MHz, $\chi_{bb}=20.7$ MHz and $\chi_{cc}=18.6$ MHz, while for $PF_{2}^{37}Cl$, $\chi_{aa}= 0-1.31$ MHz, $\chi_{bb}= 16.6$ MHz and $\chi_{cc}= 14.5$ MHz

Development of a thin section device for space exploration: Rock cutting mechanism

We have developed a rock cutting mechanism for in situ planetary exploration based on abrasive diamond impregnated wire. Performance characteristics of the rock cutter, including cutting rate on several rock types, cutting surface lifetime, and cut rock surface finish are presented. The rock cutter was developed as part of a broader effort to develop an in situ automated rock thin section (IS-ARTS) instrument. The objective of IS-ARTS was to develop an instrument capable of producing petrographic rock thin sections on a planetary science spacecraft. The rock cutting mechanism may also be useful to other planetary science missions with in situ instruments in which sub-sampling and rock surface preparation are necessary. © 2012 COSPAR. Published by Elsevier Ltd. All rights reserved

Development of a thin section device for space exploration: Overview and system performance estimates

In this paper we present a conceptual design of a spaceborne instrument for the in situ production of rock thin sections on planetary surfaces. The in situ Automated Rock Thin Section Instrument (IS-ARTS) conceptual design demonstrates that the in situ production of thin sections on a planetary body is a plausible new instrument capability for future planetary exploration. Thin section analysis would reduce much ambiguity in the geological history of a sampled site that is present with instruments currently flown. The technical challenge of producing a thin section device compatible with the spacecraft environment is formidable and has been thought too technically difficult to be practical. Terrestrial thin section preparation requires a skilled petrographist, several preparation instruments that individually exceed typical spacecraft mass and power limits, and consumable materials that are not easily compatible with spaceflight. In two companion papers we present research and development work used to constrain the capabilities of IS-ARTS in the technical space compatible with the spacecraft environment. For the design configuration shown we conclude that a device can be constructed that is capable of 50 sample preparations over a 2 year lifespan with mass, power, and volume constraints compatible with current landed Mars mission configurations. The technical requirements of IS-ARTS (mass, power and number of samples produced) depend strongly on the sample mechanical properties, sample processing rate, the sample size and number of samples to be produced. © 2012 COSPAR. Published by Elsevier Ltd. All rights reserved

Development of a thin section device for space exploration: Overview and system performance estimates

In this paper we present a conceptual design of a spaceborne instrument for the in situ production of rock thin sections on planetary surfaces. The in situ Automated Rock Thin Section Instrument (IS-ARTS) conceptual design demonstrates that the in situ production of thin sections on a planetary body is a plausible new instrument capability for future planetary exploration. Thin section analysis would reduce much ambiguity in the geological history of a sampled site that is present with instruments currently flown. The technical challenge of producing a thin section device compatible with the spacecraft environment is formidable and has been thought too technically difficult to be practical. Terrestrial thin section preparation requires a skilled petrographist, several preparation instruments that individually exceed typical spacecraft mass and power limits, and consumable materials that are not easily compatible with spaceflight. In two companion papers we present research and development work used to constrain the capabilities of IS-ARTS in the technical space compatible with the spacecraft environment. For the design configuration shown we conclude that a device can be constructed that is capable of 50 sample preparations over a 2 year lifespan with mass, power, and volume constraints compatible with current landed Mars mission configurations. The technical requirements of IS-ARTS (mass, power and number of samples produced) depend strongly on the sample mechanical properties, sample processing rate, the sample size and number of samples to be produced. © 2012 COSPAR. Published by Elsevier Ltd. All rights reserved