Molecular Masking of Synthetic Immunomodulator Evokes Antitumor Immunity With Reduced Immune Tolerance and Systemic Toxicity by Temporal Activity Recovery and Sustained Stimulation.

Activation of the innate immune system counteracts tumor-induced immunosuppression. Hence, small molecule-based toll-like receptor 7/8 agonists (TLR7/8a), which can modulate immunosuppression in the tumor microenvironment along with the activation of innate immunity, are emerging as essential components of cancer immunotherapy. However, the clinical application of synthetic TLR7/8a therapies is limited by systemic immune-associated toxicity and immune tolerance induced by uncontrolled stimulatory activities and repeated treatments. To address these limitations, a dynamic immunomodulation strategy incorporating masking and temporal recovery of the activity of TLR7/8a through prodrug-like TLR7/8a (pro-TLR7/8a) at the molecular level and a sustained and controlled release of active TLR7/8a from nanoliposome (pro-TLR7/8a) (NL(pro-TLR7/8)) in a macroscale depot are designed. Immunization with cationic NL(pro-TLR7/8) and anionic antigens triggers robust activation of innate immune cells as well as antigen-specific T cell responses, eliciting reprogramming of immunosuppressive cells into tumor-suppressive cells, with decreased systemic adverse effects and immune tolerance. Combination treatment with NL(pro-TLR7/8a) and immune checkpoint inhibitors (anti-CTLA-4 plus anti-PD-L1) or nanoliposomes (Doxorubicin) has synergistic effects on antitumor immunity in various tumor models. The concept of pro-TLR7/8a suggested herein may facilitate the advancement of small-molecule-based immunomodulators for clinical translation and safe and effective cancer immunotherapy.

A nanoadjuvant that dynamically coordinates innate immune stimuli activation enhances cancer immunotherapy and reduces immune cell exhaustion.

Although conventional innate immune stimuli contribute to immune activation, they induce exhausted immune cells, resulting in suboptimal cancer immunotherapy. Here we suggest a kinetically activating nanoadjuvant (K-nanoadjuvant) that can dynamically integrate two waves of innate immune stimuli, resulting in effective antitumour immunity without immune cell exhaustion. The combinatorial code of K-nanoadjuvant is optimized in terms of the order, duration and time window between spatiotemporally activating Toll-like receptor 7/8 agonist and other Toll-like receptor agonists. K-nanoadjuvant induces effector/non-exhausted dendritic cells that programme the magnitude and persistence of interleukin-12 secretion, generate effector/non-exhausted CD8+ T cells, and activate natural killer cells. Treatment with K-nanoadjuvant as a monotherapy or in combination therapy with anti-PD-L1 or liposomes (doxorubicin) results in strong antitumour immunity in murine models, with minimal systemic toxicity, providing a strategy for synchronous and dynamic tailoring of innate immunity for enhanced cancer immunotherapy.

Overcoming Chemoimmunotherapy-Induced Immunosuppression by Assemblable and Depot Forming Immune Modulating Nanosuspension.

The deficiency of antigen-specific T cells and the induction of various treatment-induced immunosuppressions still limits the clinical benefit of cancer immunotherapy. Although the chemo-immunotherapy adjuvanted with Toll-like receptor 7/8 agonist (TLR 7/8a) induces immunogenic cell death (ICD) and in situ vaccination effect, indoleamine 2,3-dioxygenase (IDO) is also significantly increased in the tumor microenvironment (TME) and tumor-draining lymph node (TDLN), which offsets the activated antitumor immunity. To address the treatment-induced immunosuppression, an assemblable immune modulating suspension (AIMS) containing ICD inducer (paclitaxel) and supra-adjuvant (immune booster; R848 as a TLR 7/8a, immunosuppression reliever; epacadostat as an IDO inhibitor) is suggested and shows that it increases cytotoxic T lymphocytes and relieves the IDO-related immunosuppression (TGF-ß, IL-10, myeloid-derived suppressor cells, and regulatory T cells) in both TME and TDLN, by the formation of in situ depot in tumor bed as well as by the efficient migration into TDLN. Local administration of AIMS increases T cell infiltration in both local and distant tumors and significantly inhibits the metastasis of tumors to the lung. Reverting treatment-induced secondary immunosuppression and reshaping "cold tumor" into "hot tumor" by AIMS also increases the response rate of immune checkpoint blockade therapy, which promises a new nanotheranostic strategy in cancer immunotherapy.

Chemical Strategies to Enhance the Therapeutic Efficacy of Toll-like Receptor Agonist Based Cancer Immunotherapy.

Recent developments in the fields of biomedical chemistry and immune bioengineering have enabled innovative therapeutic approaches that can enhance the efficacy, accuracy, and safety of cancer immunotherapy. Among the numerous strategies utilized in cancer immunotherapy, Toll-like receptor (TLR) agonist-based approaches have been studied for a long time since they trigger the innate immune system and generate antigen-specific T cell responses to fight against tumors. In addition to these immunostimulatory functions, TLR agonists also contribute to the reprogramming of immune suppressive tumor microenvironments. Although TLR agonists are now being intensively studied in clinical trials due to their substantial immunomodulatory properties, they still show a low therapeutic index. Nonspecific and random stimulation of various immune cells produces excess levels of proinflammatory cytokines, resulting in cytokine storms and chronic diseases. Therefore, the development of chemical strategies to enhance the therapeutic efficacy as well as the safety of TLR agonist-based immunotherapy is essential and in high demand.In this Account, we summarize and discuss recent developments in biomedical chemistry and bioengineering techniques for the immunomodulation of TLR agonists that have addressed the limitations in current cancer immunotherapy. Immunomodulation of TLR agonists can be classified into two different approaches: (1) molecular modulation via chemical structure modification and (2) macroscopic modulation via an engineered drug delivery system. In molecular modulation, based on prodrug and antedrug principles, activity is modulated (active or inactive) through immolative chemical linkers that can respond to extrinsic or intrinsic biological stimulation and the plasmatic environment, respectively. To increase the effectiveness of TLR agonists as immunostimulatory agents, researchers have conjugated TLR agonists with other immunotherapeutic moieties (antigen, antibody, other TLR agonist, etc.). For macroscopic modulation, bioengineering of delivery carriers differing in size or with albumin hitchhiking moieties has been utilized to increase the efficiency of the targeting of these carriers to secondary lymphoid organs (lymph nodes (LNs) and spleen). The conjugation of specific targeting ligands and incorporation of stimulus-triggering moieties can promote the delivery of TLR agonists into specific cells or intracellular compartments. Implantable porous scaffolds for specific immune cell recruitment and in situ depot-forming gel systems for controlled release of immunomodulatory drugs can increase the therapeutic efficacy of TLR agonists while reducing systemic toxicity. Taken together, these findings show that well-designed and precisely controlled chemical strategies for the immunomodulation of TLR agonists at both the molecular and macroscopic levels are expected to play key roles in improving the therapeutic efficacy of cancer immunotherapy while minimizing immune-related toxicity.

A three-dimensional hyaluronic acid-based niche enhances the therapeutic efficacy of human natural killer cell-based cancer immunotherapy.

Adoptive transfer of natural killer (NK) cells is becoming one of the most important parts of cancer immunotherapy. However, recent accomplishments have focused on the improvement of the targeting effects based on the engineering of chimeric antigen receptors (CARs) on cell surfaces. Despite the large quantity of therapeutic cells required for clinical applications, the technology for ex vivo expansion is not well developed. Herein, a three-dimensional (3D) engineered hyaluronic acid-based niche for cell expansion (3D-ENHANCE) is introduced. Compared with the conventional two-dimensional (2D) method, NK-92 cell lines and human EGFR-specific (CAR)-NK cells cultured in 3D-ENHANCE yield favorable mRNA expressions, elevated cytokine release, upregulated proliferative and tumor-lytic abilities, and result in enhanced antitumor efficacy. Furthermore, controllable degradation rates can be realized by tuning the formulation of 3D-ENHANCE so that it can be applied as an implantable cell reservoir at surgical sites. In vivo results with the incompletely resected MDA-MB-231 model confirm that the peri-operative implantation of 3D-ENHANCE prevents the relapse and metastases after surgery. Overall, 3D-ENHANCE presents an effective cytokine-free niche for ex vivo expansion and postsurgical treatment that enhances the low-therapeutic efficacy of human NK cells.

Lyophilizable and Multifaceted Toll-like Receptor 7/8 Agonist-Loaded Nanoemulsion for the Reprogramming of Tumor Microenvironments and Enhanced Cancer Immunotherapy.

The low therapeutic efficacy of current cancer immunotherapy is related to nonimmunogenic and immunosuppressive tumor microenvironments (TMEs). To overcome these limitations, both the immune priming of antitumoral lymphocytes and the reprogramming of immunosuppressive factors in TMEs are essential. Here, we suggest a nanoemulsion (NE)-based immunotherapeutic platform that can not only modulate tumor-induced suppression but also induce an effective cell-mediated immune response for T cell proliferation. Multifunctional NEs can be fabricated by integrating the efficacy of NEs as delivery systems and the multifaceted immunomodulation characteristics (i.e., immunostimulation and reprogramming of immunosuppression) of small molecule-based Toll-like receptor 7/8 agonists. Local in situ vaccination of melanoma and cervical tumor models with tumor antigens (protein and peptide) adjuvanted with NE loaded with TLR7/8 agonists [NE (TLR7/8a)] induced the recruitment and activation of innate immune cells, infiltration of lymphocytes, and polarization of tumor-associated M2 macrophages, which resulted in inhibition of tumor growth and prolonged survival in both primary and rechallenged tumor models. Antibody-depletion experiments also suggested that macrophages, type I IFN (IFN-α and IFN-ß), CD8+ T cells, and NK1.1+ cells contributed to the antitumor effect of NE (TLR7/8a). The combination of antitumoral lymphocytes and reprogramming of immunosuppressive TMEs induced by NE (TLR7/8a) treatment evoked a synergistic antitumor immune response with immune checkpoint blockade therapy (anti-PD-1 and anti-PD-L1).

Syringeable immunotherapeutic nanogel reshapes tumor microenvironment and prevents tumor metastasis and recurrence.

The low response rate of current cancer immunotherapy suggests the presence of few antigen-specific T cells and a high number of immunosuppressive factors in tumor microenvironment (TME). Here, we develop a syringeable immunomodulatory multidomain nanogel (iGel) that overcomes the limitation by reprogramming of the pro-tumoral TME to antitumoral immune niches. Local and extended release of immunomodulatory drugs from iGel deplete immunosuppressive cells, while inducing immunogenic cell death and increased immunogenicity. When iGel is applied as a local postsurgical treatment, both systemic antitumor immunity and a memory T cell response are generated, and the recurrence and metastasis of tumors to lungs and other organs are significantly inhibited. Reshaping of the TME using iGel also reverts non-responding groups to checkpoint blockade therapies into responding groups. The iGel is expected as an immunotherapeutic platform that can reshape immunosuppressive TMEs and synergize cancer immunotherapy with checkpoint therapies, with minimized systemic toxicity.

Analysis of structure and P-c-T curve of hydrogenated Ti53Zr27xNi20Pd(x) quasicrystals.

The potential application of TiZrNi quasicrystals was evaluated by measuring the pressure-composition-temperature curves after replacing Zr by Pd to the limit maintaining the host structure for the samples made with Ti53Zr27(-x)Ni20Pd(x), where 0 < or = x < or = 8. The results of X-ray diffraction data revealed that the samples keep the pure quasicrystal structure to the maximum value of x = 8. All diffracted peaks uniformly shifted to the low angle of 2 theta in X-ray diffraction pattern suggesting that hydrogen atoms homogeneously diffuse in the quasicrystals and uniformly expand the quasilattice constants without modification of the structure. After hydrogenation at elevated temperature, the quasi-lattice constants increased from 5.12 to 5.34 angstroms for the samples made with x = 0 without appearing an impurity phase. When Zr was replaced by 8 at.% of Pd, the equilibrium vapor pressures significantly increased to 3.41 from 0.41 Torr at 300 degrees C although the total amount of hydrogen decreased as increasing Pd concentration. These results demonstrate that Pd will play a critical role in application for the TiZrNi quasicrystals as hydrogen storage materials.

The effect of thermal annealing on pentacene thin film transistor with micro contact printing.

We used micro contact printing (micro-CP) to fabricate inverted coplanar pentacene thin film transistors (TFTs) with 1-microm channels. The patterning of micro-scale source/drain electrodes without etch process was successfully achieved using Polydimethylsiloxane (PDMS) elastomer stamp. We used the Ag nano particle ink as an electrode material, and the sheet resistance and surface roughness of the Ag electrodes were effectively reduced with the 2-step thermal annealing on a hotplate, which improved the mobility, the on-off ratio, and the subthreshold slope (SS) of the pentacene TFTs. In addition, the device annealing on a hotplate in a N2 atmosphere for 30 sec can enhance the off-current and the mobility properties of OTFTs without damaging the pentacene thin films and increase the adhesion between pentacene and dielectric layer (SiO2), which was investigated with the pentacene films phase change of the XRD spectrum after device annealing.

Novel palladium germanide schottky contact for high performance schottky barrier ge MOSFETs and characterization of its leakage current mechanism.

The leakage current mechanism of Palladium (Pd) germanide Schottky contact on n-type Ge-on-Si substrate is analyzed in depth. The electric field dependent analysis shows that the dominant leakage current mechanism is the Poole-Frenkel emission due to the existence of deep level traps in the depletion region of the Pd germanide/n-type Ge Schottky diode. The analysis of the dependence of leakage current on temperature also shows that the Poole-Frenkel emission and generation current are the dominant components below 100 degrees C and that the Schottky emission related to thermionic emission of majority carriers over a potential barrier is the main cause of this dominance at high temperature region.

Formation of metal and dielectric liners using a solution process for deep trench capacitors.

We demonstrated the feasibility of metal and dielectric liners using a solution process for deep trench capacitor application. The deep Si trench via with size of 10.3 microm and depth of 71 microm were fabricated by Bosch process in deep reactive ion etch (DRIE) system. The aspect ratio was about 7. Then, nano-Ag ink and poly(4-vinylphenol) (PVPh) were used to form metal and dielectric liners, respectively. The thicknesses of the Ag and PVPh liners were about 144 and 830 nm, respectively. When the curing temperature of Ag film increased from 120 to 150 degrees C, the sheet resistance decreased rapidly from 2.47 to 0.72 Omega/sq and then slightly decreased to 0.6 Omega/sq with further increasing the curing temperature beyond 150 degrees C. The proposed liner formation method using solution process is a simple and cost effective process for the high capacity of deep trench capacitor.

Thermal endurance and microstructural evolution of PtGe for high-performance nano-scale Ge-on-Si MOSFETS.

The thermal endurance and microstructural evolution of Ni-germanide (NiGe) and Pt-germanide (PtGe) on a Ge-on-Si substrate were compared in this paper. In case of the Ni/TiN structure, the sheet resistance exhibited a stable RTP window of 350 to 600 degrees C, while that of the Pt/TiN structure showed more stable characteristics up to 700 degrees C. Furthermore, after post-germanidation annealing, NiGe exhibited the formation of islands due to the severe agglomeration as well as a prominent grain boundary grooving, which accounts for the sharp increase of the sheet resistance from 550 degrees C, whereas PtGe showed a smooth and continuous surface morphological stability without signs of agglomeration even up to 600 degrees C. Although about two times higher resistivity (31.5 micro ohms-cm) and greater Ge consumption (3.27 nm) were shown, PtGe showed more stable sheet resistance, better surface and interface morphological stability and a wider thermal processing window above 100 degrees C than NiGe. Therefore, PtGe is more suitable for the germanided shallow source/drain for nano-scale Ge MOSFETs than NiGe.

Improvement of heavy dopant doped Ni-silicide using ytterbium interlayer for nano-scale MOSFETS with an ultra shallow junction.

In this paper, a novel Ni silicide with Yb interlayer (Yb/Ni/TiN) on a boron cluster (B18H22) implanted source/drain junction is proposed for the first time, and its thermal stability characteristics are analyzed in depth. The proposed Ni-silicide exhibits a wider RTP temperature window for uniform sheet resistance, surface roughness and better thermal stability than the conventional structure (Ni/TiN). In addition, the cross-sectional profile of the proposed Ni-silicide showed less agglomeration despite the high temperature post-silicidation annealing, and it can be said that the proposed structure was little dependence on the temperature post-silicidation annealing. The improvement of Ni silicide properties is analyzed and found to be due to the formation of the rare earth metal--NiSi (YbNi2Si2), whose peaks were confirmed by XRD. The junction leakage current of the p + -n junction with Yb/Ni/TiN and B18H22 implantation is smaller than that with Ni/TiN by almost one order of magnitude as well as improving the thermal stability of ultra shallow junction.

Study of surface-modified PVP gate dielectric in organic thin film transistors with the nano-particle silver ink source/drain electrode.

We have fabricated the flexible pentacene based organic thin film transistors (OTFTs) with formulated poly[4-vinylphenol] (PVP) gate dielectrics treated by CF4/O2 plasma on poly[ethersulfones] (PES) substrate. The solution of gate dielectrics is made by adding methylated poly[melamine-co-formaldehyde] (MMF) to PVP. The PVP gate dielectric layer was cross linked at 90 degrees under UV ozone exposure. Source/drain electrodes are formed by micro contact printing (MCP) method using nano particle silver ink for the purposes of low cost and high throughput. The optimized OTFT shows the device performance with field effect mobility of the 0.88 cm2/V s, subthreshold slope of 2.2 V/decade, and on/off current ratios of 1.8 x 10(-6) at -40 V gate bias. We found that hydrophobic PVP gate dielectric surface can influence on the initial film morphologies of pentacene making dense, which is more important for high performance OTFTs than large grain size. Moreover, hydrophobic gate dielelctric surface reduces voids and -OH groups that interrupt the carrier transport in OTFTs.

Maskless laser direct patterning of PEDOT/PSS layer for soluble process organic thin film transistor.

In this work, we have fabricated TIPS-pentacene TFTs with conductive polymer (3,4-ethylenedioxythiophene):poly(4-stylenesulfonate) (PEDOT:PSS) source/drain electrodes which is patterned by maskless laser direct patterning (LDP). The 5-microm resolution of source and drain patterns with PEDOT:PSS were clearly defined. Furthermore, the OTFTs with 10-microm channel length were successfully achieved by exposing the focused Neodymium:Yttrium Aluminum Garnet (Nd:YAG) laser beam onto the spin-coated PEDOT:PSS films and developing with deionized water. The electrical performance of the TIPS-pentacene TFTs with PEDOT:PSS source/drain electrodes were improved with decrease of the sheet resistance of PEDOT:PSS films when the PEDOT:PSS films were annealed at the temperature above 200 degrees C.

Sub-50 nm template fabrications for nanoimprint lithography using hydrogen silsesquioxane and silicon nitride.

The sub-50 nm templates are successfully fabricated using hydrogen silsesquioxane (HSQ) and silicon nitride on silicon substrate. The HSQ template is directly patterned by e-beam direct writing. The cured HSQ pattern is used for the template of nanoimprint process. The silicon nitride template is reactively ion etched by ZEP resist mask pattern which is prepared by e-beam direct writing using ZEP resist. The line widths of HSQ templates and ZEP patterns after developments are between 22-30 nm and 24-30 nm, respectively. The line width of silicon nitride templates without performing descum is same as that of the ZEP pattern but shows a rough surface. When plasma descum was performed before RIE, the line width of silicon nitride templates increased from 27 nm to 35 nm and has a clean surface. The HSQ template fabrication results in this study will be promise for sub-nm imprint process.