Comparison of the optical quality vision between real post-LASIK myopic laser surgery and the simulated implantation of a phakic IOL in low myopia.

A phakic intraocular lens (PIOL) of - 4.5 D was characterized from its wavefront aberration profile. A preclinical study was conducted using pre- and post-surgery data from four patients that had undergone myopic laser refractive surgery. All these patients would have needed a PIOL of - 4.5 D. Pre-surgery data were used to simulate the effect of a PIOL implantation. Post myopic refractive surgery data were used to calculate the post-LASIK eye model. Modulation transfer function (MTF), point spread function (PSF) and simulation of optotypes vision were obtained and compared. The PIOL did not worsen the optical quality of the eyes evaluated. High order Aberrations were always higher in the post-LASIK eye model. Optics quality trended to be better in PIOL implantation than post-LASIK surgery as pupil size increased.

MET-Induced CD73 Restrains STING-Mediated Immunogenicity of EGFR-Mutant Lung Cancer.

Immunotherapy has shown limited efficacy in patients with EGFR-mutated lung cancer. Efforts to enhance the immunogenicity of EGFR-mutated lung cancer have been unsuccessful to date. Here, we discover that MET amplification, the most common mechanism of resistance to third-generation EGFR tyrosine kinase inhibitors (TKI), activates tumor cell STING, an emerging determinant of cancer immunogenicity (1). However, STING activation was restrained by ectonucleosidase CD73, which is induced in MET-amplified, EGFR-TKI-resistant cells. Systematic genomic analyses and cell line studies confirmed upregulation of CD73 in MET-amplified and MET-activated lung cancer contexts, which depends on coinduction of FOSL1. Pemetrexed (PEM), which is commonly used following EGFR-TKI treatment failure, was identified as an effective potentiator of STING-dependent TBK1-IRF3-STAT1 signaling in MET-amplified, EGFR-TKI-resistant cells. However, PEM treatment also induced adenosine production, which inhibited T-cell responsiveness. In an allogenic humanized mouse model, CD73 deletion enhanced immunogenicity of MET-amplified, EGFR-TKI-resistant cells, and PEM treatment promoted robust responses regardless of CD73 status. Using a physiologic antigen recognition model, inactivation of CD73 significantly increased antigen-specific CD8+ T-cell immunogenicity following PEM treatment. These data reveal that combined PEM and CD73 inhibition can co-opt tumor cell STING induction in TKI-resistant EGFR-mutated lung cancers and promote immunogenicity. SIGNIFICANCE: MET amplification upregulates CD73 to suppress tumor cell STING induction and T-cell responsiveness in TKI-resistant, EGFR-mutated lung cancer, identifying a strategy to enhance immunogenicity and improve treatment.

Activation of Tumor-Cell STING Primes NK-Cell Therapy.

Activation of the stimulator of interferon genes (STING) pathway promotes antitumor immunity but STING agonists have yet to achieve clinical success. Increased understanding of the mechanism of action of STING agonists in human tumors is key to developing therapeutic combinations that activate effective innate antitumor immunity. Here, we report that malignant pleural mesothelioma cells robustly express STING and are responsive to STING agonist treatment ex vivo. Using dynamic single-cell RNA sequencing of explants treated with a STING agonist, we observed CXCR3 chemokine activation primarily in tumor cells and cancer-associated fibroblasts, as well as T-cell cytotoxicity. In contrast, primary natural killer (NK) cells resisted STING agonist-induced cytotoxicity. STING agonists enhanced migration and killing of NK cells and mesothelin-targeted chimeric antigen receptor (CAR)-NK cells, improving therapeutic activity in patient-derived organotypic tumor spheroids. These studies reveal the fundamental importance of using human tumor samples to assess innate and cellular immune therapies. By functionally profiling mesothelioma tumor explants with elevated STING expression in tumor cells, we uncovered distinct consequences of STING agonist treatment in humans that support testing combining STING agonists with NK and CAR-NK cell therapies.

Analysis and comparison of monofocal, extended depth of focus and trifocal intraocular lens profiles.

To test the feasibility of using profilometers to extract information about IOL surfaces design. A standard monofocal IOL (Tecnis 1), a monofocal IOL that provided some depth of focus (Eyhance), an extended depth of focus IOL based on refractive optics (Mini Well) and a trifocal IOL based on diffractive optics were used in this study (Tecnis Synergy). The surface topography of the IOLs was measured by using a multimode optical profilometer. Posterior surface of Tecnis 1 IOL was spherical and the anterior surface aspherical. In the Eyhance IOL, posterior surface was spherical and anterior surface did not fit to any of our reference surfaces, indicating a higher order aspheric surface design. In the Mini Well Ready IOL, a best-fit sphere surface was obtained for the second surface and a high order aspherical surface design was deduced for the first surface. The anterior surface of the Synergy IOL was aspherical and the base curve of the diffractive structure fitted very well to a spherical surface. To consider an aspheric surface as possible best-fit surface provided more information than if only best-fit spherical surface was considered. The high order aspheric surface designs employed in the IOLs studied presented differences, regarding best-fit asphere surface, higher than 1 micron. These differences were correlated with the generation of spherical aberration complex profiles (with Zernike terms higher than 4th order) and with the production of distinct amounts of depth of focus. This method was also useful to deduce the base curve of diffractive surfaces.

Dynamic single-cell RNA sequencing identifies immunotherapy persister cells following PD-1 blockade.

Resistance to oncogene-targeted therapies involves discrete drug-tolerant persister cells, originally discovered through in vitro assays. Whether a similar phenomenon limits efficacy of programmed cell death 1 (PD-1) blockade is poorly understood. Here, we performed dynamic single-cell RNA-Seq of murine organotypic tumor spheroids undergoing PD-1 blockade, identifying a discrete subpopulation of immunotherapy persister cells (IPCs) that resisted CD8+ T cell-mediated killing. These cells expressed Snai1 and stem cell antigen 1 (Sca-1) and exhibited hybrid epithelial-mesenchymal features characteristic of a stem cell-like state. IPCs were expanded by IL-6 but were vulnerable to TNF-α-induced cytotoxicity, relying on baculoviral IAP repeat-containing protein 2 (Birc2) and Birc3 as survival factors. Combining PD-1 blockade with Birc2/3 antagonism in mice reduced IPCs and enhanced tumor cell killing in vivo, resulting in durable responsiveness that matched TNF cytotoxicity thresholds in vitro. Together, these data demonstrate the power of high-resolution functional ex vivo profiling to uncover fundamental mechanisms of immune escape from durable anti-PD-1 responses, while identifying IPCs as a cancer cell subpopulation targetable by specific therapeutic combinations.

New method to improve the quality of vision in cataractous keratoconus eyes.

To analyze using optical simulations if the proper use of a segmented intraocular lens (IOL) can improve the visual outcomes compared to the implantation of a spherical monofocal IOL. The wavefront profile of the Mplus (Oculentis) and a monofocal IOLs with the phase transformation introduced by each IOL were calculated using a Hartmann-Shack wavefront sensor. In addition, the wavefront profile of schematic eye models of various keratoconus conditions was obtained and was propagated to the IOLs. The optical performance of such combination was obtained after combining ray tracing and Fourier optics. A pre-clinical validation was also evaluated incorporating clinical data from three different keratoconus eyes of three patients. The implantation of the Mplus IOL can compensate or reduce the overall coma of the eye with keratoconus improving the quality of vision compared with a spherical monofocal IOL due to lower displacements of the retinal image or tilting in keratoconus. All theoretical simulations were confirmed afterwards by mean of a preclinical validation. The use of a standard toric segmented IOL with a proper orientation and selection of the addition can improve the optical quality of the keratoconus eye compared to the use of a monofocal spherical IOL.

Suppression of Myeloid Cell Arginase Activity leads to Therapeutic Response in a NSCLC Mouse Model by Activating Anti-Tumor Immunity.

BACKGROUND: Tumor orchestrated metabolic changes in the microenvironment limit generation of anti-tumor immune responses. Availability of arginine, a semi-essential amino acid, is critical for lymphocyte proliferation and function. Levels of arginine are regulated by the enzymes arginase 1,2 and nitric oxide synthase (NOS). However, the role of arginase activity in lung tumor maintenance has not been investigated in clinically relevant orthotopic tumor models. METHODS: RNA sequencing (RNA-seq) of sorted cell populations from mouse lung adenocarcinomas derived from immunocompetent genetically engineered mouse models (GEMM)s was performed. To complement mouse studies, a patient tissue microarray consisting of 150 lung adenocarcinomas, 103 squamous tumors, and 54 matched normal tissue were stained for arginase, CD3, and CD66b by multiplex immunohistochemistry. Efficacy of a novel arginase inhibitor compound 9 in reversing arginase mediated T cell suppression was determined in splenocyte ex vivo assays. Additionally, the anti-tumor activity of this compound was determined in vitro and in an autochthonous immunocompetent KrasG12D GEMM of lung adenocarcinoma model. RESULTS: Analysis of RNA-seq of sorted myeloid cells suggested that arginase expression is elevated in myeloid cells in the tumor as compared to the normal lung tissue. Accordingly, in the patient samples arginase 1 expression was mainly localized in the granulocytic myeloid cells and significantly elevated in both lung adenocarcinoma and squamous tumors as compared to the controls. Our ex vivo analysis demonstrated that myeloid derived suppressor cell (MDSC)s cause T cell suppression by arginine depletion, and suppression of arginase activity by a novel ARG1/2 inhibitor, compound 9, led to restoration of T cell function by increasing arginine. Treatment of KrasG12D GEMM of lung cancer model with compound 9 led to a significant tumor regression associated with increased T cell numbers and function, while it had no activity across several murine and human non-small cell (NSCLC) lung cancer lines in vitro. CONCLUSIONS: We show that arginase expression is elevated in mouse and patient lung tumors. In a KRASG12D GEMM arginase inhibition diminished growth of established tumors. Our data suggest arginase as an immunomodulatory target that should further be investigated in lung tumors with high arginase activity.

Simulation of the Effect of Different Presbyopia-Correcting Intraocular Lenses With Eyes With Previous Laser Refractive Surgery.

PURPOSE: To simulate the optical performance of three presbyopia-correcting intraocular lenses (IOLs) implanted in eyes with previous laser refractive surgery. METHODS: A simulation of the through-focus modulation transfer function (MTF) was performed for three presbyopia-correcting IOLs (Mplus, Oculentis GmbH, Berlin, Germany; Symfony, Johnson & Johnson Vision, Santa Ana, CA; and Mini Well, SIFI S.p.A., Lavinaio, Italy) in one eye with previous myopic LASIK and another with hyperopic LASIK. Real topographic data and the wavefront aberration profile of each IOL obtained with a Hartmann-Shack sensor were used. RESULTS: In the eye with myopic LASIK, all IOLs lost optical quality at near and intermediate distances for 4- and 4.7-mm pupil size. For 3-mm pupil size, the Mini Well IOL showed the best intermediate and near MTF and maintained the far focus independently of the pupil. In the eye with hyperopic LASIK, the Mini Well IOL showed an intermediate, distance, and -4.00-diopter (D) foci for all pupils. The Symfony IOL showed a depth of focus at far and intermediate distance for 3-mm and a focus at -2.50 D in the rest. The Mplus showed a focus of -4.50 and -3.00 D for the 3- and 4-mm pupil, respectively. CONCLUSIONS: The Mini Well and Symfony IOLs seem to work better than the Mplus IOL in eyes with previous myopic LASIK. With previous hyperopic LASIK, the Mini Well IOL seems to be able to provide acceptable near, intermediate, and far foci for all pupil sizes. These findings should be confirmed in future clinical studies. [J Refract Surg. 2018;34(4):222-227.].

In Vitro Aberrometric Assessment of a Multifocal Intraocular Lens and Two Extended Depth of Focus IOLs.

PURPOSE: To analyze the "in vitro" aberrometric pattern of a refractive IOL and two extended depth of focus IOLs. METHODS: A special optical bench with a Shack-Hartmann wavefront sensor (SH) was designed for the measurement. Three presbyopia correction IOLs were analyzed: Mini WELL (MW), TECNIS Symfony ZXR00 (SYM), and Lentis Mplus X LS-313 MF30 (MP). Three different pupil sizes were used for the comparison: 3, 4, and 4.7 mm. RESULTS: MW generated negative primary and positive secondary spherical aberrations (SA) for the apertures of 3 mm (-0.13 and +0.12 µm), 4 mm (-0.12 and +0.08 µm), and 4.7 mm (-0.11 and +0.08 µm), while the SYM only generated negative primary SA for 4 and 4.7 mm apertures (-0.12 µm and -0.20 µm, resp.). The MP induced coma and trefoil for all pupils and showed significant HOAs for apertures of 4 and 4.7 mm. CONCLUSIONS: In an optical bench, the MW induces negative primary and positive secondary SA for all pupils. The SYM aberrations seem to be pupil dependent; it does not produce negative primary SA for 3 mm but increases for higher pupils. Meanwhile, the HOAs for the MW and SYM were not significant. The MP showed in all cases the highest HOAs.

The Potent ALK Inhibitor Brigatinib (AP26113) Overcomes Mechanisms of Resistance to First- and Second-Generation ALK Inhibitors in Preclinical Models.

PURPOSE: Non-small cell lung cancers (NSCLCs) harboring ALK gene rearrangements (ALK+) typically become resistant to the first-generation anaplastic lymphoma kinase (ALK) tyrosine kinase inhibitor (TKI) crizotinib through development of secondary resistance mutations in ALK or disease progression in the brain. Mutations that confer resistance to second-generation ALK TKIs ceritinib and alectinib have also been identified. Here, we report the structure and first comprehensive preclinical evaluation of the next-generation ALK TKI brigatinib. EXPERIMENTAL DESIGN: A kinase screen was performed to evaluate the selectivity profile of brigatinib. The cellular and in vivo activities of ALK TKIs were compared using engineered and cancer-derived cell lines. The brigatinib-ALK co-structure was determined. RESULTS: Brigatinib potently inhibits ALK and ROS1, with a high degree of selectivity over more than 250 kinases. Across a panel of ALK+ cell lines, brigatinib inhibited native ALK (IC50, 10 nmol/L) with 12-fold greater potency than crizotinib. Superior efficacy of brigatinib was also observed in mice with ALK+ tumors implanted subcutaneously or intracranially. Brigatinib maintained substantial activity against all 17 secondary ALK mutants tested in cellular assays and exhibited a superior inhibitory profile compared with crizotinib, ceritinib, and alectinib at clinically achievable concentrations. Brigatinib was the only TKI to maintain substantial activity against the most recalcitrant ALK resistance mutation, G1202R. The unique, potent, and pan-ALK mutant activity of brigatinib could be rationalized by structural analyses. CONCLUSIONS: Brigatinib is a highly potent and selective ALK inhibitor. These findings provide the molecular basis for the promising activity being observed in ALK+, crizotinib-resistant patients with NSCLC being treated with brigatinib in clinical trials. Clin Cancer Res; 22(22); 5527-38. ©2016 AACR.

Novel Small Molecule Inhibitors of Choline Kinase Identified by Fragment-Based Drug Discovery.

Choline kinase α (ChoKα) is an enzyme involved in the synthesis of phospholipids and thereby plays key roles in regulation of cell proliferation, oncogenic transformation, and human carcinogenesis. Since several inhibitors of ChoKα display antiproliferative activity in both cellular and animal models, this novel oncogene has recently gained interest as a promising small molecule target for cancer therapy. Here we summarize our efforts to further validate ChoKα as an oncogenic target and explore the activity of novel small molecule inhibitors of ChoKα. Starting from weakly binding fragments, we describe a structure based lead discovery approach, which resulted in novel highly potent inhibitors of ChoKα. In cancer cell lines, our lead compounds exhibit a dose-dependent decrease of phosphocholine, inhibition of cell growth, and induction of apoptosis at low micromolar concentrations. The druglike lead series presented here is optimizable for improvements in cellular potency, drug target residence time, and pharmacokinetic parameters. These inhibitors may be utilized not only to further validate ChoKα as antioncogenic target but also as novel chemical matter that may lead to antitumor agents that specifically interfere with cancer cell metabolism.

Ultrathin high-index metasurfaces for shaping focused beams.

The volume size of a converging wave, which plays a relevant role in image resolution, is governed by the wavelength of the radiation and the numerical aperture (NA) of the wavefront. We designed an ultrathin (λ/8 width) curved metasurface that is able to transform a focused field into a high-NA optical architecture, thus boosting the transverse and (mainly) on-axis resolution. The elements of the metasurface are metal-insulator subwavelength gratings exhibiting extreme anisotropy with ultrahigh index of refraction for TM polarization. Our results can be applied to nanolithography and optical microscopy.

Engineered surface waves in hyperbolic metamaterials.

We analyzed surface-wave propagation that takes place at the boundary between a semi-infinite dielectric and a multilayered metamaterial, the latter with indefinite permittivity and cut normally to the layers. Known hyperbolization of the dispersion curve is discussed within distinct spectral regimes, including the role of the surrounding material. Hybridization of surface waves enable tighter confinement near the interface in comparison with pure-TM surface-plasmon polaritons. We demonstrate that the effective-medium approach deviates severely in practical implementations. By using the finite-element method, we predict the existence of long-range oblique surface waves.

Fragment growing and linking lead to novel nanomolar lactate dehydrogenase inhibitors.

Lactate dehydrogenase A (LDH-A) catalyzes the interconversion of lactate and pyruvate in the glycolysis pathway. Cancer cells rely heavily on glycolysis instead of oxidative phosphorylation to generate ATP, a phenomenon known as the Warburg effect. The inhibition of LDH-A by small molecules is therefore of interest for potential cancer treatments. We describe the identification and optimization of LDH-A inhibitors by fragment-based drug discovery. We applied ligand based NMR screening to identify low affinity fragments binding to LDH-A. The dissociation constants (K(d)) and enzyme inhibition (IC(50)) of fragment hits were measured by surface plasmon resonance (SPR) and enzyme assays, respectively. The binding modes of selected fragments were investigated by X-ray crystallography. Fragment growing and linking, followed by chemical optimization, resulted in nanomolar LDH-A inhibitors that demonstrated stoichiometric binding to LDH-A. Selected molecules inhibited lactate production in cells, suggesting target-specific inhibition in cancer cell lines.

Left-handed metamaterial coatings for subwavelength-resolution imaging.

We report on a procedure to improve the resolution of far-field imaging by using a neighboring high-index medium that is coated with a left-handed metamaterial. The resulting plot can also exhibit an enhanced transmission by considering proper conditions to retract backscattering. Based on negative refraction, geometrical aberrations are considered in detail since they may cause a great impact in this sort of diffraction-unlimited imaging by reducing its resolution power. We employ a standard aberration analysis to refine the asymmetric configuration of metamaterial superlenses. We demonstrate that low-order centrosymmetric aberrations can be fully corrected for a given object plane. For subwavelength-resolution imaging, however, high-order aberrations become of relevance, which may be balanced with defocus. Not only the point spread function but also numerical simulations based on the finite-element method support our theoretical analysis, and subwavelength resolution is verified in the image plane.

Considerations on the electromagnetic flow in Airy beams based on the Gouy phase.

We reexamine the Gouy phase in ballistic Airy beams (AiBs). A physical interpretation of our analysis is derived in terms of the local phase velocity and the Poynting vector streamlines. Recent experiments employing AiBs are consistent with our results. We provide an approach which potentially applies to any finite-energy paraxial wave field that lacks a beam axis.

Algorithm for correcting the keratometric estimation error in normal eyes..

PURPOSE: To obtain an accurate algorithm for calculating the keratometric index that minimizes the errors in the calculation of corneal power assuming only a single corneal surface in the range of corneal curvatures of the normal population. METHODS: Corneal power was calculated by using the classical keratometric index and also by using the Gaussian equation. Differences between types of calculation of corneal power were determined and modeled by regression analysis. RESULTS: We proposed two options for the selection of the most appropriate keratometric index (n(k)) value for each specific case. First was the use of specific linear equations (depending on the ratio of the anterior to the posterior curvature, k ratio) according to the value of the central radius of curvature of the anterior corneal surface (r(1c)) in 0.1 mm steps and the theoretical eye model considered. The second was the use of a general simplified equation only requiring r(1c) (Gullstrand eye model, n(k) = -0.0064286r(1c) + 1.37688; Le Grand eye model, n(k) = -0.0063804r(1c) + 1.37806). CONCLUSIONS: The generalization of the keratometric index (n(k)) value is not an appropriate approximation for the estimation of the corneal power and it can lead to significant errors. We proposed a new algorithm depending on r(1c), with a maximal associated error in the calculation of the corneal power of 0.5 D and without requiring knowledge of the posterior corneal curvature.

Crizotinib-resistant mutants of EML4-ALK identified through an accelerated mutagenesis screen.

Activating gene rearrangements of anaplastic lymphoma kinase (ALK) have been identified as driver mutations in non-small-cell lung cancer, inflammatory myofibroblastic tumors, and other cancers. Crizotinib, a dual MET/ALK inhibitor, has demonstrated promising clinical activity in patients with non-small-cell lung cancer and inflammatory myofibroblastic tumors harboring ALK translocations. Inhibitors of driver kinases often elicit kinase domain mutations that confer resistance, and such mutations have been successfully predicted using in vitro mutagenesis screens. Here, this approach was used to discover an extensive set of ALK mutations that can confer resistance to crizotinib. Mutations at 16 residues were identified, structurally clustered into five regions around the kinase active site, which conferred varying degrees of resistance. The screen successfully predicted the L1196M, C1156Y, and F1174L mutations, recently identified in crizotinib-resistant patients. In separate studies, we demonstrated that crizotinib has relatively modest potency in ALK-positive non-small-cell lung cancer cell lines. A more potent ALK inhibitor, TAE684, maintained substantial activity against mutations that conferred resistance to crizotinib. Our study identifies multiple novel mutations in ALK that may confer clinical resistance to crizotinib, suggests that crizotinib's narrow selectivity window may underlie its susceptibility to such resistance and demonstrates that a more potent ALK inhibitor may be effective at overcoming resistance.

Nondiffracting Bessel plasmons.

We report on the existence of nondiffracting Bessel surface plasmon polaritons (SPPs), advancing at either superluminal or subluminal phase velocities. These wave fields feature deep subwavelength FWHM, but are supported by high-order homogeneous SPPs of a metal/dielectric (MD) superlattice. The beam axis can be relocated to any MD interface, by interfering multiple converging SPPs with controlled phase matching. Dissipative effects in metals lead to a diffraction-free regime that is limited by the energy attenuation length. However, the ultra-localization of the diffracted wave field might still be maintained by more than one order of magnitude.

Three-dimensional point spread function and generalized amplitude transfer function of near-field flat lenses.

We derive a nonsingular, polarization-dependent, 3D impulse response that provides unambiguously the wave field scattered by a negative-refractive-index layered lens and distributed in its image volume. By means of a 3D Fourier transform, we introduce the generalized amplitude transfer function in order to gain a deep insight into the resolution power of the optical element. In the near-field regime, fine details containing some depth information may be transmitted through the lens. We show that metamaterials with moderate absorption are appropriate for subwavelength resolution keeping a limited degree of depth discrimination.