Wavelength-Sensitive Superconducting Single-Photon Detectors on Thin Film Lithium Niobate Waveguides.

Lithium niobate, because of its nonlinear and electro-optical properties, is one of the materials of choice for photonic applications. The development of nanostructuring capabilities of thin film lithium niobate (TFLN) permits fabrication of small footprint, low-loss optical circuits. With the recent implementation of on-chip single-photon detectors, this architecture is among the most promising for realizing on-chip quantum optics experiments. In this Letter, we report on the implementation of superconducting nanowire single-photon detectors (SNSPDs) based on NbTiN on 300 nm thick TFLN ridge nano-waveguides. We demonstrate a waveguide-integrated wavelength meter based on the photon energy dependence of the superconducting detectors. The device operates at the telecom C- and L-bands and has a footprint smaller than 300 × 180 µm2 and critical currents between ∼12 and ∼14 µA, which ensures operation with minimum heat dissipation. Our results hold promise for future densely packed on-chip wavelength-multiplexed quantum communication systems.

Biomarker Discovery in Rare Malignancies: Development of a miRNA Signature for RDEB-cSCC.

Machine learning has been proven to be a powerful tool in the identification of diagnostic tumor biomarkers but is often impeded in rare cancers due to small patient numbers. In patients suffering from recessive dystrophic epidermolysis bullosa (RDEB), early-in-life development of particularly aggressive cutaneous squamous-cell carcinomas (cSCCs) represents a major threat and timely detection is crucial to facilitate prompt tumor excision. As miRNAs have been shown to hold great potential as liquid biopsy markers, we characterized miRNA signatures derived from cultured primary cells specific for the potential detection of tumors in RDEB patients. To address the limitation in RDEB-sample accessibility, we analyzed the similarity of RDEB miRNA profiles with other tumor entities derived from the Cancer Genome Atlas (TCGA) repository. Due to the similarity in miRNA expression with RDEB-SCC, we used HN-SCC data to train a tumor prediction model. Three models with varying complexity using 33, 10 and 3 miRNAs were derived from the elastic net logistic regression model. The predictive performance of all three models was determined on an independent HN-SCC test dataset (AUC-ROC: 100%, 83% and 96%), as well as on cell-based RDEB miRNA-Seq data (AUC-ROC: 100%, 100% and 91%). In addition, the ability of the models to predict tumor samples based on RDEB exosomes (AUC-ROC: 100%, 93% and 100%) demonstrated the potential feasibility in a clinical setting. Our results support the feasibility of this approach to identify a diagnostic miRNA signature, by exploiting publicly available data and will lay the base for an improvement of early RDEB-SCC detection.

5'RNA Trans-Splicing Repair of COL7A1 Mutant Transcripts in Epidermolysis Bullosa.

Mutations within the COL7A1 gene underlie the inherited recessive subtype of the blistering skin disease dystrophic epidermolysis bullosa (RDEB). Although gene replacement approaches for genodermatoses are clinically advanced, their implementation for RDEB is challenging and requires endogenous regulation of transgene expression. Thus, we are using spliceosome-mediated RNA trans-splicing (SMaRT) to repair mutations in COL7A1 at the mRNA level. Here, we demonstrate the capability of a COL7A1-specific RNA trans-splicing molecule (RTM), initially selected using a fluorescence-based screening procedure, to accurately replace COL7A1 exons 1 to 64 in an endogenous setting. Retroviral RTM transduction into patient-derived, immortalized keratinocytes resulted in an increase in wild-type transcript and protein levels, respectively. Furthermore, we revealed accurate deposition of recovered type VII collagen protein within the basement membrane zone of expanded skin equivalents using immunofluorescence staining. In summary, we showed for the first time the potential of endogenous 5' trans-splicing to correct pathogenic mutations within the COL7A1 gene. Therefore, we consider 5' RNA trans-splicing a suitable tool to beneficially modulate the RDEB-phenotype, thus targeting an urgent need of this patient population.

Strain-Controlled Quantum Dot Fine Structure for Entangled Photon Generation at 1550 nm.

Entangled photon generation at 1550 nm in the telecom C-band is of critical importance as it enables the realization of quantum communication protocols over long distance using deployed telecommunication infrastructure. InAs epitaxial quantum dots have recently enabled on-demand generation of entangled photons in this wavelength range. However, time-dependent state evolution, caused by the fine-structure splitting, currently limits the fidelity to a specific entangled state. Here, we show fine-structure suppression for InAs quantum dots using micromachined piezoelectric actuators and demonstrate generation of highly entangled photons at 1550 nm. At the lowest fine-structure setting, we obtain a maximum fidelity of 90.0 ± 2.7% (concurrence of 87.5 ± 3.1%). The concurrence remains high also for moderate (weak) temporal filtering, with values close to 80% (50%), corresponding to 30% (80%) of collected photons, respectively. The presented fine-structure control opens the way for exploiting entangled photons from quantum dots in fiber-based quantum communication protocols.

On-Demand Generation of Entangled Photon Pairs in the Telecom C-Band with InAs Quantum Dots.

Entangled photons are an integral part in quantum optics experiments and a key resource in quantum imaging, quantum communication, and photonic quantum information processing. Making this resource available on-demand has been an ongoing scientific challenge with enormous progress in recent years. Of particular interest is the potential to transmit quantum information over long distances, making photons the only reliable flying qubit. Entangled photons at the telecom C-band could be directly launched into single-mode optical fibers, enabling worldwide quantum communication via existing telecommunication infrastructure. However, the on-demand generation of entangled photons at this desired wavelength window has been elusive. Here, we show a photon pair generation efficiency of 69.9 ± 3.6% in the telecom C-band by an InAs/GaAs semiconductor quantum dot on a metamorphic buffer layer. Using a robust phonon-assisted two-photon excitation scheme we measure a maximum concurrence of 91.4 ± 3.8% and a peak fidelity to the Φ+ state of 95.2 ± 1.1%, verifying on-demand generation of strongly entangled photon pairs and marking an important milestone for interfacing quantum light sources with our classical fiber networks.

Personalized Development of Antisense Oligonucleotides for Exon Skipping Restores Type XVII Collagen Expression in Junctional Epidermolysis Bullosa.

Intermediate junctional epidermolysis bullosa caused by mutations in the COL17A1 gene is characterized by the frequent development of blisters and erosions on the skin and mucous membranes. The rarity of the disease and the heterogeneity of the underlying mutations renders therapy developments challenging. However, the high number of short in-frame exons facilitates the use of antisense oligonucleotides (AON) to restore collagen 17 (C17) expression by inducing exon skipping. In a personalized approach, we designed and tested three AONs in combination with a cationic liposomal carrier for their ability to induce skipping of COL17A1 exon 7 in 2D culture and in 3D skin equivalents. We show that AON-induced exon skipping excludes the targeted exon from pre-mRNA processing, which restores the reading frame, leading to the expression of a slightly truncated protein. Furthermore, the expression and correct deposition of C17 at the dermal-epidermal junction indicates its functionality. Thus, we assume AON-mediated exon skipping to be a promising tool for the treatment of junctional epidermolysis bullosa, particularly applicable in a personalized manner for rare genotypes.

Proximitized Josephson junctions in highly-doped InAs nanowires robust to optical illumination.

We have studied the effects of optical-frequency light on proximitized InAs/Al Josephson junctions based on highly n-doped InAs nanowires at varying incident photon flux and at three different photon wavelengths. The experimentally obtained IV curves were modeled using a resistively shunted junction model which takes scattering at the contact interfaces into account. Despite the fact that the InAs weak link is photosensitive, the Josephson junctions were found to be surprisingly robust, interacting with the incident radiation only through heating, whereas above the critical current our devices showed non-thermal effects resulting from photon exposure. Our work indicates that Josephson junctions based on highly-doped InAs nanowires can be integrated in close proximity to photonic circuits. The results also suggest that such junctions can be used for optical-frequency photon detection through thermal processes by measuring a shift in critical current.

Crux of Using the Cascaded Emission of a Three-Level Quantum Ladder System to Generate Indistinguishable Photons.

We investigate the degree of indistinguishability of cascaded photons emitted from a three-level quantum ladder system; in our case the biexciton-exciton cascade of semiconductor quantum dots. For the three-level quantum ladder system we theoretically demonstrate that the indistinguishability is inherently limited for both emitted photons and determined by the ratio of the lifetimes of the excited and intermediate states. We experimentally confirm this finding by comparing the quantum interference visibility of noncascaded emission and cascaded emission from the same semiconductor quantum dot. Quantum optical simulations produce very good agreement with the measurements and allow us to explore a large parameter space. Based on our model, we propose photonic structures to optimize the lifetime ratio and overcome the limited indistinguishability of cascaded photon emission from a three-level quantum ladder system.

Origin of Antibunching in Resonance Fluorescence.

Resonance fluorescence has played a major role in quantum optics with predictions and later experimental confirmation of nonclassical features of its emitted light such as antibunching or squeezing. In the Rayleigh regime where most of the light originates from the scattering of photons with subnatural linewidth, antibunching would appear to coexist with sharp spectral lines. Here, we demonstrate that this simultaneous observation of subnatural linewidth and antibunching is not possible with simple resonant excitation. Using an epitaxial quantum dot for the two-level system, we independently confirm the single-photon character and subnatural linewidth by demonstrating antibunching in a Hanbury Brown and Twiss type setup and using high-resolution spectroscopy, respectively. However, when filtering the coherently scattered photons with filter bandwidths on the order of the homogeneous linewidth of the excited state of the two-level system, the antibunching dip vanishes in the correlation measurement. Our observation is explained by antibunching originating from photon-interferences between the coherent scattering and a weak incoherent signal in a skewed squeezed state. This prefigures schemes to achieve simultaneous subnatural linewidth and antibunched emission.

A cancer stem cell-like phenotype is associated with miR-10b expression in aggressive squamous cell carcinomas.

BACKGROUND: Cutaneous squamous cell carcinomas (cSCC) are the primary cause of premature deaths in patients suffering from the rare skin-fragility disorder recessive dystrophic epidermolysis bullosa (RDEB), which is in marked contrast to the rarely metastasizing nature of these carcinomas in the general population. This remarkable difference is attributed to the frequent development of chronic wounds caused by impaired skin integrity. However, the specific molecular and cellular changes to malignancy, and whether there are common players in different types of aggressive cSCCs, remain relatively undefined. METHODS: MiRNA expression profiling was performed across various cell types isolated from skin and cSCCs. Microarray results were confirmed by qPCR and by an optimized in situ hybridization protocol. Functional impact of overexpression or knock-out of a dysregulated miRNA was assessed in migration and 3D-spheroid assays. Sample-matched transcriptome data was generated to support the identification of disease relevant miRNA targets. RESULTS: Several miRNAs were identified as dysregulated in cSCCs compared to control skin. These included the metastasis-linked miR-10b, which was significantly upregulated in primary cell cultures and in archival biopsies. At the functional level, overexpression of miR-10b conferred the stem cell-characteristic of 3D-spheroid formation capacity to keratinocytes. Analysis of miR-10b downstream effects identified a novel putative target of miR-10b, the actin- and tubulin cytoskeleton-associated protein DIAPH2. CONCLUSION: The discovery that miR-10b mediates an aspect of cancer stemness - that of enhanced tumor cell adhesion, known to facilitate metastatic colonization - provides an important avenue for future development of novel therapies targeting this metastasis-linked miRNA.

GaAs Quantum Dot in a Parabolic Microcavity Tuned to 87Rb D1.

We develop a structure to efficiently extract photons emitted by a GaAs quantum dot tuned to rubidium. For this, we employ a broadband microcavity with a curved gold backside mirror that we fabricate by a combination of photoresist reflow, dry reactive ion etching in an inductively coupled plasma, and selective wet chemical etching. Precise reflow and etching control allows us to achieve a parabolic backside mirror with a short focal distance of 265 nm. The fabricated structures yield a predicted (measured) collection efficiency of 63% (12%), an improvement by more than 1 order of magnitude compared to unprocessed samples. We then integrate our quantum dot parabolic microcavities onto a piezoelectric substrate capable of inducing a large in-plane biaxial strain. With this approach, we tune the emission wavelength by 0.5 nm/kV, in a dynamic, reversible, and linear way, to the rubidium D1 line (795 nm).

Optimizing the stoichiometry of ultrathin NbTiN films for high-performance superconducting nanowire single-photon detectors.

The requirements in quantum optics experiments for high single-photon detection efficiency, low timing jitter, low dark count rate and short dead time have been fulfilled with the development of superconducting nanowire single-photon detectors. Although they offer a detection efficiency above 90%, achieving a high time resolution in devices made of amorphous materials is a challenge, particularly at temperatures above 0.8 K. Devices made from niobium nitride and niobium titanium nitride allow us to reach the best timing jitter but, in turn, have stronger requirements in terms of film quality to achieve a high efficiency. Here we take advantage of the flexibility of reactive co-sputter deposition to tailor the composition of NbxTi1-xN superconducting films and show that a Nb fraction of x = 0.62 allows for the fabrication of detectors from films as thick as 9 nm and covering an active area of 20 µm, with a wide detection saturation plateau at telecom wavelengths and in particular at 1550 nm. This is a signature of an internal detection efficiency saturation, achieved while maintaining the high time resolution associated with NbTiN and operation at 2.5K. With our optimized recipe, we reliably fabricated detectors with high critical current densities reaching a saturation plateau at 1550 nm with 80% system detection efficiency and with a FWHM timing jitter as low as 19.5 ps.

Gene Editing-Mediated Disruption of Epidermolytic Ichthyosis-Associated KRT10 Alleles Restores Filament Stability in Keratinocytes.

Epidermolytic ichthyosis is a skin fragility disorder caused by dominant-negative mutations in KRT1 or KRT10. No definitive restorative therapies exist that target these genetic faults. Gene editing can be used to efficiently introduce frameshift mutations to inactivate mutant genes. This can be applied to counter the effect of dominantly inherited diseases such as epidermolytic ichthyosis. In this study, we used transcription activator-like effector nuclease technology, to disrupt disease-causing mutant KRT10 alleles in an ex vivo cellular approach, with the intent of developing a therapy for patients with epidermolytic ichthyosis. A transcription activator-like effector nuclease was designed to specifically target a region of KRT10, upstream of a premature termination codon known to induce a genetic knockout. This proved highly efficient at gene disruption in a patient-derived keratinocyte cell line. In addition, analysis for off-target effects indicated no promiscuous gene editing-mediated disruption. Reversion of the keratin intermediate filament fragility phenotype associated with epidermolytic ichthyosis was observed by the immunofluorescence analysis of correctly gene-edited single-cell clones. This was in concurrence with immunofluorescence and ultrastructure analysis of murine xenograft models. The efficiency of this approach was subsequently confirmed in primary patient keratinocytes. Our data demonstrate the feasibility of an ex vivo gene-editing therapy for more than 95.6% of dominant KRT10 mutations.

Resonance Fluorescence of GaAs Quantum Dots with Near-Unity Photon Indistinguishability.

Photonic quantum technologies call for scalable quantum light sources that can be integrated, while providing the end user with single and entangled photons on demand. One promising candidate is strain free GaAs/AlGaAs quantum dots obtained by aluminum droplet etching. Such quantum dots exhibit ultra low multi-photon probability and an unprecedented degree of photon pair entanglement. However, different to commonly studied InGaAs/GaAs quantum dots obtained by the Stranski-Krastanow mode, photons with a near-unity indistinguishability from these quantum emitters have proven to be elusive so far. Here, we show on-demand generation of near-unity indistinguishable photons from these quantum emitters by exploring pulsed resonance fluorescence. Given the short intrinsic lifetime of excitons and trions confined in the GaAs quantum dots, we show single photon indistinguishability with a raw visibility of [Formula: see text], without the need for Purcell enhancement. Our results represent a milestone in the advance of GaAs quantum dots by demonstrating the final missing property standing in the way of using these emitters as a key component in quantum communication applications, e.g., as quantum light sources for quantum repeater architectures.

Strain-Tunable Quantum Integrated Photonics.

Semiconductor quantum dots are crucial parts of the photonic quantum technology toolbox because they show excellent single-photon emission properties in addition to their potential as solid-state qubits. Recently, there has been an increasing effort to deterministically integrate single semiconductor quantum dots into complex photonic circuits. Despite rapid progress in the field, it remains challenging to manipulate the optical properties of waveguide-integrated quantum emitters in a deterministic, reversible, and nonintrusive manner. Here we demonstrate a new class of hybrid quantum photonic circuits combining III-V semiconductors, silicon nitride, and piezoelectric crystals. Using a combination of bottom-up, top-down, and nanomanipulation techniques, we realize strain tuning of a selected, waveguide-integrated, quantum emitter and a planar integrated optical resonator. Our findings are an important step toward realizing reconfigurable quantum-integrated photonics, with full control over the quantum sources and the photonic circuit.

Basal pharmacokinetic parameters of topically applied diacerein in pediatric patients with generalized severe epidermolysis bullosa simplex.

Generalized severe epidermolysis bullosa simplex (EBS-gen sev) is caused by mutations within either the KRT5 or KRT14 gene, phenotypically resulting in blistering and wounding of the skin and mucous membranes after minor mechanical friction. In a clinical phase 2/3 trial, diacerein has recently been shown to significantly reduce blister numbers upon topical application. In this study we addressed basic pharmacokinetic parameters of locally applied diacerein in vitro and in vivo. Ex vivo experiments using a Franz diffusion cell confirmed the uptake and bio-transformation of diacerein to rhein in a porcine skin model. Rhein, the active metabolite of diacerein, was also detected in both urine and serum samples of two EBS-gen sev patients who topically applied a 1% diacerein ointment over a period of 4 weeks. The accumulated systemic levels of rhein in EBS-gen sev patients were lower than reported levels after oral application. These preliminary findings point towards the uptake and prolonged persistance of diacerein / rhein within the intended target organ - the skin. Further, they imply an acceptable safety profile at the systemic level. TRIAL REGISTRATION: DRKS. DRKS00005412 . Registered 6 November 2013.

Diacerein orphan drug development for epidermolysis bullosa simplex: A phase 2/3 randomized, placebo-controlled, double-blind clinical trial.

BACKGROUND: Epidermolysis bullosa simplex (EBS) is a rare genetic, blistering skin disease for which there is no cure. Treatments that address the pathophysiology of EBS are needed. OBJECTIVE: Compare the impact of 1% diacerein cream with placebo in reducing the number of blisters in EBS. METHODS: In a randomized, placebo-controlled, phase 2/3 trial we used a 1% diacerein topical formulation to treat defined skin areas in 17 patients. In a 2-period crossover trial, patients were randomized to either placebo or diacerein for a 4-week treatment and a 3-month follow-up in period 1. After a washout, patients were crossed over during period 2. The prespecified primary end point was the proportion of patients with a reduction of number of blisters by more than 40% from baseline in selected areas over the treatment episode. RESULTS: Of the patients receiving diacerein, 86% in episode 1 and 37.5% in episode 2 met the primary end point (vs 14% and 17% with placebo, respectively). This effect was still significant after the follow-up. Changes in absolute blister numbers were significant for the diacerein group only. No adverse effects were observed. LIMITATIONS: Low patient numbers and no invasive data acquisition because of clinical burden in children. CONCLUSION: This trial provides evidence of the impact of 1% diacerein cream in the treatment of EBS.

Threonine 150 Phosphorylation of Keratin 5 Is Linked to Epidermolysis Bullosa Simplex and Regulates Filament Assembly and Cell Viability.

A characteristic feature of the skin blistering disease epidermolysis bullosa simplex is keratin filament (KF) network collapse caused by aggregation of the basal epidermal keratin type II (KtyII) K5 and its type I partner keratin 14 (K14). Here, we examine the role of keratin phosphorylation in KF network rearrangement and cellular functions. We detect phosphorylation of the K5 head domain residue T150 in cytoplasmic epidermolysis bullosa simplex granules containing R125C K14 mutants. Expression of phosphomimetic T150D K5 mutants results in impaired KF formation in keratinocytes. The phenotype is enhanced upon combination with other phosphomimetic K5 head domain mutations. Remarkably, introduction of T150D K5 mutants into KtyII-lacking (KtyII-/-) keratinocytes prevents keratin network formation altogether. In contrast, phosphorylation-deficient T150A K5 leads to KFs with reduced branching and turnover. Assembly of T150D K5 is arrested at the heterotetramer stage coinciding with increased heat shock protein association. Finally, reduced cell viability and elevated response to stressors is noted in T150 mutant cells. Taken together, our findings identify T150 K5 phosphorylation as an important determinant of KF network formation and function with a possible role in epidermolysis bullosa simplex pathogenesis.

Cut and Paste: Efficient Homology-Directed Repair of a Dominant Negative KRT14 Mutation via CRISPR/Cas9 Nickases.

With the ability to induce rapid and efficient repair of disease-causing mutations, CRISPR/Cas9 technology is ideally suited for gene therapy approaches for recessively and dominantly inherited monogenic disorders. In this study, we have corrected a causal hotspot mutation in exon 6 of the keratin 14 gene (KRT14) that results in generalized severe epidermolysis bullosa simplex (EBS-gen sev), using a double-nicking strategy targeting intron 7, followed by homology-directed repair (HDR). Co-delivery into EBS keratinocytes of a Cas9 D10A nickase (Cas9n), a predicted single guide RNA pair specific for intron 7, and a minicircle donor vector harboring the homology donor template resulted in a recombination efficiency of >30% and correction of the mutant KRT14 allele. Phenotypic correction of EBS-gen sev keratinocytes was demonstrated by immunofluorescence analysis, revealing the absence of disease-associated K14 aggregates within the cytoplasm. We achieved a promising safety profile for the CRISPR/Cas9 double-nicking approach, with no detectable off-target activity for a set of predicted off-target genes as confirmed by next generation sequencing. In conclusion, we demonstrate a highly efficient and specific gene-editing approach for KRT14, offering a causal treatment option for EBS.