Protein Signature Differentiating Neutrophils and Myeloid-Derived Suppressor Cells Determined Using a Human Isogenic Cell Line Model and Protein Profiling.

Myeloid-derived suppressor cells (MDSCs) play an essential role in suppressing the antitumor activity of T lymphocytes in solid tumors, thus representing an attractive therapeutic target to enhance the efficacy of immunotherapy. However, the differences in protein expression between MDSCs and their physiological counterparts, particularly polymorphonuclear neutrophils (PMNs), remain inadequately characterized, making the specific identification and targeting of MDSCs difficult. PMNs and PMN-MDSCs share markers such as CD11b+CD14-CD15+/CD66b+, and some MDSC-enriched markers are emerging, such as LOX-1 and CD84. More proteomics studies are needed to identify the signature and markers for MDSCs. Recently, we reported the induced differentiation of isogenic PMNs or MDSCs (referred to as iPMNs and iMDSCs, respectively) from the human promyelocytic cell line HL60. Here, we profiled the global proteomics and membrane proteomics of these cells with quantitative mass spectrometry, which identified a 41-protein signature ("cluster 6") that was upregulated in iMDSCs compared with HL60 and iPMN. We further integrated our cell line-based proteomics data with a published proteomics dataset of normal human primary monocytes and monocyte-derived MDSCs induced by cancer-associated fibroblasts. The analysis identified a 38-protein signature that exhibits an upregulated expression pattern in MDSCs compared with normal monocytes or PMNs. These signatures may provide a hypothesis-generating platform to identify protein biomarkers that phenotypically distinguish MDSCs from their healthy counterparts, as well as potential therapeutic targets that impair MDSCs without harming normal myeloid cells.

Ketogenic Diet Alters the Epigenetic and Immune Landscape of Prostate Cancer to Overcome Resistance to Immune Checkpoint Blockade Therapy.

Resistance to immune checkpoint blockade (ICB) therapy represents a formidable clinical challenge limiting the efficacy of immunotherapy. In particular, prostate cancer poses a challenge for ICB therapy due to its immunosuppressive features. A ketogenic diet (KD) has been reported to enhance response to ICB therapy in some other cancer models. However, adverse effects associated with continuous KD were also observed, demanding better mechanistic understanding and optimized regimens for using KD as an immunotherapy sensitizer. In this study, we established a series of ICB-resistant prostate cancer cell lines and developed a highly effective strategy of combining anti-PD1 and anti-CTLA4 antibodies with histone deacetylase inhibitor (HDACi) vorinostat, a cyclic KD (CKD), or dietary supplementation of the ketone body ß-hydroxybutyrate (BHB), which is an endogenous HDACi. CKD and BHB supplementation each delayed prostate cancer tumor growth as monotherapy, and both BHB and adaptive immunity were required for the antitumor activity of CKD. Single-cell transcriptomic and proteomic profiling revealed that HDACi and ketogenesis enhanced ICB efficacy through both cancer cell-intrinsic mechanisms, including upregulation of MHC class I molecules, and -extrinsic mechanisms, such as CD8+ T-cell chemoattraction, M1/M2 macrophage rebalancing, monocyte differentiation toward antigen-presenting cells, and diminished neutrophil infiltration. Overall, these findings illuminate a potential clinical path of using HDACi and optimized KD regimens to enhance ICB therapy for prostate cancer. SIGNIFICANCE: Optimized cyclic ketogenic diet and 1,3-butanediol supplementation regimens enhance the efficacy of immune checkpoint blockade in prostate cancer through epigenetic and immune modulations, providing dietary interventions to sensitize tumors to immunotherapy.

Controllable Circularly Polarized Luminescence with High Dissymmetry Factor via Co-Assembly of Achiral Dyes in Liquid Crystal Polymer Films.

Circularly polarized luminescence (CPL) materials are highly demanded due to their great potential in optoelectronic and chiroptical elements. However, the preparation of CPL films with high luminescence dissymmetry factors (glum ) remains a formidable task, which impedes their practical application in film-based devices. Herein, a facile strategy to prepare solid CPL film with a high glum through exogenous chiral induction and amplification of liquid crystal polymers is proposed. Amplification and reversion of the CPL appear when the films are annealed at the chiral nematic liquid crystalline temperature and the maximal glum up to 0.30 due to the enhancement of selective reflection. Thermal annealing treatment at different liquid crystalline states facilitates the formation of the chiral liquid phase and adjusts the circularly polarized emission. This work not only provides a straightforward and versatile platform to construct organic films capable of exhibiting strong circularly polarized emission but also is helpful in understanding the exact mechanism for the liquid crystal enhancement of CPL performance.

Neutrophils resist ferroptosis and promote breast cancer metastasis through aconitate decarboxylase 1.

Metastasis causes breast cancer-related mortality. Tumor-infiltrating neutrophils (TINs) inflict immunosuppression and promote metastasis. Therapeutic debilitation of TINs may enhance immunotherapy, yet it remains a challenge to identify therapeutic targets highly expressed and functionally essential in TINs but under-expressed in extra-tumoral neutrophils. Here, using single-cell RNA sequencing to compare TINs and circulating neutrophils in murine mammary tumor models, we identified aconitate decarboxylase 1 (Acod1) as the most upregulated metabolic enzyme in mouse TINs and validated high Acod1 expression in human TINs. Activated through the GM-CSF-JAK/STAT5-C/EBPß pathway, Acod1 produces itaconate, which mediates Nrf2-dependent defense against ferroptosis and upholds the persistence of TINs. Acod1 ablation abates TIN infiltration, constrains metastasis (but not primary tumors), bolsters antitumor T cell immunity, and boosts the efficacy of immune checkpoint blockade. Our findings reveal how TINs escape from ferroptosis through the Acod1-dependent immunometabolism switch and establish Acod1 as a target to offset immunosuppression and improve immunotherapy against metastasis.

Overcome Prostate Cancer Resistance to Immune Checkpoint Therapy with Ketogenic Diet-Induced Epigenetic Reprogramming.

Advanced prostate cancer (PCa) is overwhelmingly resistant to immune checkpoint blockade (ICB) therapy, representing a formidable clinical challenge. In this study, we developed a syngeneic murine PCa model with acquired ICB resistance. Using this model, synergistic efficacy was achieved by combining anti-PD1 and anti-CTLA4 antibodies with histone deacetylase inhibitor (HDACi) vorinostat, a cyclic ketogenic diet (CKD), or supplementation of ketone body ß-hydroxybutyrate (BHB, endogenous HDACi) via 1,3-butanediol-admixed food. CKD and BHB supplementation delayed PCa tumors as monotherapy, and both BHB and adaptive immunity are required for the anti-tumor activity of CKD. Single-cell transcriptomic and proteomic profiling revealed that the HDACi and ketogenesis-enhanced ICB therapy involves cancer-cell-intrinsic (upregulated MHC class I molecules) and extrinsic mechanisms (CD8 + T cell chemoattraction, M1/M2 macrophage rebalancing, monocyte differentiation toward antigen presenting cells, and diminished neutrophils). Overall, these findings underscore the potential of using HDACi and optimized KD to enhance ICB therapy for PCa.

Ovarian juvenile granulosa cell tumors with Ollier's disease in children with IDH1 gene somatic mutation.

Objective: The aim of this study was to explore the symptoms, treatment, and pathogenesis of ovarian juvenile granulosa cell tumors with Ollier's disease in children. Methods: From October 2019 to October 2020, clinical data were retrospectively analyzed for one case of ovarian juvenile granulosa cell tumors with Ollier's disease. Whole-exome sequencing and Sanger sequencing were used to detect gene mutation in ovarian tumor and chondroma tissue. NADP-dependent isocitrate dehydrogenase-1 (IDH1) and S6 ribosomal protein expression levels in cells transfected with wild-type or mutant plasmid were analyzed by Western blot. Results: The 4-year-old female showed multiple skeletal deformities, bilateral breast development with chromatosis, and vulvar discharge. Sex hormone assay suggested that estradiol and prolactin were elevated, and the x-ray of limbs suggested enchondroma. Pelvic ultrasound and abdominal CT revealed a right ovarian solid mass. Pathologic examination of the right ovarian solid mass showed a juvenile granulosa cell type. A c.394C>T (p. Arg132Cys) mutation of the IDH1 gene was detected in both the ovarian juvenile granulosa cell tumors and enchondroma. Transfection of HeLa cells with either WT or Mut plasmid caused 4.46- or 3.77-fold overexpression of IDH1 gene compared to non-transfected control cells, respectively. R132C mutation inhibited the phosphorylation of S6 ribosomal protein, which is central to the mTOR pathway. Postoperatively, estradiol and prolactin levels fell to values normal for her age and bilateral breast gradual retraction. Conclusion: The incidence of ovarian juvenile granulosa cell tumors with Ollier's disease in children may be caused by generalized mesodermal dysplasia; IDH1 gene mutation may play a facilitated role in this process. Surgical operation is the main treatment. We suggest that patients with ovarian juvenile granulosa cell tumors and Ollier's disease should undergo regular investigation.

Targeting the chromatin effector Pygo2 promotes cytotoxic T cell responses and overcomes immunotherapy resistance in prostate cancer.

The noninflamed microenvironment in prostate cancer represents a barrier to immunotherapy. Genetic alterations underlying cancer cell-intrinsic oncogenic signaling are increasingly appreciated for their role in shaping the immune landscape. Recently, we identified Pygopus 2 (PYGO2) as the driver oncogene for the amplicon at 1q21.3 in prostate cancer. Here, using transgenic mouse models of metastatic prostate adenocarcinoma, we found that Pygo2 deletion decelerated tumor progression, diminished metastases, and extended survival. Pygo2 loss augmented the activation and infiltration of cytotoxic T lymphocytes (CTLs) and sensitized tumor cells to T cell killing. Mechanistically, Pygo2 orchestrated a p53/Sp1/Kit/Ido1 signaling network to foster a microenvironment hostile to CTLs. Genetic or pharmacological inhibition of Pygo2 enhanced the antitumor efficacy of immunotherapies using immune checkpoint blockade (ICB), adoptive cell transfer, or agents inhibiting myeloid-derived suppressor cells. In human prostate cancer samples, Pygo2 expression was inversely correlated with the infiltration of CD8+ T cells. Analysis of the ICB clinical data showed association between high PYGO2 level and worse outcome. Together, our results highlight a potential path to improve immunotherapy using Pygo2-targeted therapy for advanced prostate cancer.

Cancer-cell-intrinsic mechanisms shaping the immunosuppressive landscape of prostate cancer.

Although immunotherapy has revolutionized cancer treatment and achieved remarkable success across many different cancer types, only a subset of patients shows meaningful clinical responses. In particular, advanced prostate cancer exhibits overwhelming de novo resistance to immune checkpoint blockade therapy. This is primarily due to the immunosuppressive tumor microenvironment of prostate cancer. Therefore, it is paramount to understand how prostate cancer cell-intrinsic mechanisms promote immune evasion and foster an immunosuppressive microenvironment. Here, we review recent findings that reveal the roles of the genetic alterations, androgen receptor signaling, cancer cell plasticity, and oncogenic pathways in shaping the immunosuppressive microenvironment and thereby driving immunotherapy resistance. Based on preclinical and clinical observations, a variety of therapeutic strategies are being developed that may illuminate new paths to enhance immunotherapy efficacy in prostate cancer.

Full-Color and Switchable Circularly Polarized Light from a Macroscopic Chiral Dendritic Film through a Solid-State Supramolecular Assembly.

Chiral materials displaying chirality across multiple length scales have attracted increasing interest due to their potential applications in diverse fields. Herein, we report an efficient approach for the construction of macroscopic crystal dendrites with hierarchical chirality based on an in situ solid assembly in a block copolymer film. Chiral fluorescent crystals are formed by enantiopure d-/l-dibenzoyl tartaric acid and pyrenecarboxylic acid in a poly(1,4-butadiene)-b-poly(ethylene oxide) film. The chiro-optical activity of the crystalline dendrites can be greatly amplified in the absorption and scattering regions and goes along with the dimension of dendrites. Notably, the chiral dendrites exhibited strong circularly polarized luminescence emission with a high dissymmetric factor (0.03). The enhancement of the quantum yield of the chiral film was up to 28%, which was 14 times higher that of the corresponding fluorescent molecules. The circularly polarized emission bands of the films can be fine-tuned by contriving the emissive bands of fluorescent molecules. More importantly, the chiral signals are able to be wiped when the fluorescent group photodimerizes under UV irradiation. This work provides an efficient way to develop functional materials through solid self-assembly.

Enhancing immune checkpoint blockade therapy of genitourinary malignancies by co-targeting PMN-MDSCs.

Immune checkpoint blockade (ICB) as a powerful immunotherapy has transformed cancer treatment. The application of ICB to genitourinary malignancies has generated substantial clinical benefits for patients with advanced kidney cancer or bladder cancer, yet very limited response to ICB therapy was observed from metastatic castration-resistant prostate cancer. The efficacy of ICB in rare genitourinary tumors (e.g. penile cancer) awaits results from ongoing clinical trials. A potential barrier for ICB is tumor-infiltrating polymorphonuclear myeloid-derived suppressor cells (PMN-MDSCs) with their functions and mechanisms recently revealed. Preclinical studies suggest that successful therapeutic inhibition of PMN-MDSCs synergizes effectively with ICB to eradicate ICB-refractory genitourinary malignancies.

Chondroblastoma in the Children Treated with Intralesional Curettage and Bone Grafting: Outcomes and Risk Factors for Local Recurrence.

OBJECTIVE: To review the outcomes of surgical management in the pediatric patients with extremity chondroblastoma. Especially the risk factors of recurrence and growth disorder. And discuss a potential method to decrease the rate of growth disorder by preventing the premature physeal closure. METHODS: Fifteen girls and twenty-seven boys aged from two to 14 years (mean, 11 years) with histologically proven chondroblastoma, who presented from January 2011 to June 2018 at our Hospital, were retrospectively reviewed. Clinical data, radiographic images, histological findings, treatment, functional outcomes, and local recurrence rate were analyzed. Surgical treatment included complete curettage of the tumor and the walls of the lesion followed by bone grafting, No adjuvant methods were used. Recurrence was defined as a return of symptoms and an expansion radiolucency at the operated site. It was confirmed by the histopathological analysis. When recurrence was diagnosed, the medical data were analyzed to detect the effect of different factors on local recurrence. Functional outcome was measured according to Sailhan's functional criteria, designed to provide a standardized method of assessing pediatric chondroblastoma patient postoperatively. RESULTS: The proximal part of the femur was the most frequently involved site. All the patients had at least 24 months of follow-up; mean duration was 30 months (range, 24-60 months). The local recurrence rate was 9.5%. Three resolved after repeat surgeries without further recurrence, one had a second recurrence and received another more aggressive curettage. Local recurrence of chondroblastoma was associated with age (P < 0.05), while not associated with sex, tumor location, the radiological character of the lesion or the grafting method (P > 0.05). No pulmonary metastasis was noted at latest follow-up. Five patients suffered from premature closure of physis due to physis injury. Thirty-one patients (73.8%) had a good outcome, and all returned to normal unrestricted activities. Six patients (14.3%) had a fair outcome due to occasional pain, asymmetric range of motion, or radiographic joint changes without arthritis. And five patients (11.9%) had a poor outcome because of chronic pain, loss of joint motion impairing normal life activities, or a limb-length discrepancy and limp. CONCLUSIONS: Aggressive curettage and bone grafting resulted in local control and good outcomes in most pediatric patients. Being less than 12 years of age was the risk factor for recurrence. For those growing patients, premature physeal closure was observed after the curettage, interpositional technique with PMMA would be a good choice for prevention.

Reversible Micrometer-Scale Spiral Self-Assembly in Liquid Crystalline Block Copolymer Film with Controllable Chiral Response.

The spiral is a fundamental structure in nature and spiral structures with controllable handedness are of increasing interest in the design of new chiroptical materials. In this study, micrometer-scale spiral structures with reversible chirality were fabricated based on the assembly of a liquid crystalline block copolymer film assisted by enantiopure tartaric acid. Mechanistic insight revealed that the formation of the spiral structures was closely related to the liquid crystalline properties of the major phase of block copolymer under the action of chiral tartaric acid. The chiral spiral structures with controllable handedness were easily erased under ultraviolet light irradiation and restored via thermal annealing. This facile thermal treatment method provides guidance for fabrication of chiral micrometer-scale spiral structures with adjustable chiral properties.

Spiranthes sinensis-Inspired Circular Polarized Luminescence in a Solid Block Copolymer Film with a Controllable Helix.

Chiral materials with circular polarized luminescence (CPL) have attracted much interest because of their extensive optical information and remarkable sensitivity. Inspired by the helical template in Spiranthes sinensis, we propose here a general and flexible method for fabricating solid CPL materials using a block copolymer-formed helix as a template. A chiral arrangement of various nonchiral fluorescent molecules was obtained in the block copolymer-based hybrid film. An excimer chiralty rule was found for the CPL emission of nonchiral fluorescent molecules: a right-handed helix induced left-handed CPL emission and a left-handed helix induced right-handed CPL emission. A dissipative particle dynamics simulation showed that such an antihelical effect is related to the length between the adjacent interacting points of nonchiral fluorescent molecules along the helical structure. Furthermore, the fluorescent films had a high dissymmetric factor for CPL emission, and thus, the films provide a general and flexible platform for designing and preparing advanced functional chiroptical materials.

Effective combinatorial immunotherapy for penile squamous cell carcinoma.

Penile squamous cell carcinoma (PSCC) accounts for over 95% of penile malignancies and causes significant mortality and morbidity in developing countries. Molecular mechanisms and therapies of PSCC are understudied, owing to scarcity of laboratory models. Herein, we describe a genetically engineered mouse model of PSCC, by co-deletion of Smad4 and Apc in the androgen-responsive epithelium of the penis. Mouse PSCC fosters an immunosuppressive microenvironment with myeloid-derived suppressor cells (MDSCs) as a dominant population. Preclinical trials in the model demonstrate synergistic efficacy of immune checkpoint blockade with the MDSC-diminishing drugs cabozantinib or celecoxib. A critical clinical problem of PSCC is chemoresistance to cisplatin, which is induced by Pten deficiency on the backdrop of Smad4/Apc co-deletion. Drug screen studies informed by targeted proteomics identify a few potential therapeutic strategies for PSCC. Our studies have established what we believe to be essential resources for studying PSCC biology and developing therapeutic strategies.

Limited internal fixation combined with a joint-spanning external fixator in the treatment of Midfoot injuries in children.

The aim of this study was to explore the clinical features and surgical treatment methods of unstable midfoot injuries in children. Eleven children with severe unstable midfoot injuries admitted to Jishuitan Hospital, Beijing, from June 2009 to October 2016 were enrolled, including seven patients with Lisfranc injuries and four patients with Chopart injuries. All Lisfranc injuries had radiographic data from the healthy sides, and radiographs of the affected sides showed that all injured Lisfranc joints separated more than 3 mm compared with the healthy sides. The treatment methods employed a joint-spanning external fixator to distract and maintain the length of the medial and lateral columns, combined with joint-preserving trans-joint locking plate fixation or trans-articular cannulated screw and Kirschner wire fixation that passed through the joint. The patients were followed up for 53.7 months (17-110 months). The average operation time was 95.1 minutes, and the average intraoperative blood loss was 83.3 ml. Nine of the 11 patients were treated with an external fixator to distract and fix the medial or lateral column. Bone healing was achieved in all patients, and none of the patients complained of chronic pain in the midfoot. Flatfoot and valgus deformity were corrected after the surgery in the child with old fracture, and the pain in the calcaneocuboid joint disappeared. The average American Orthopedic Foot and Ankle Society mid-foot score at the last follow-up was 93.4, of which nine cases were greater than 90 and two cases were between 75 and 89. Children's midfoot injury is characterized by fracture-dislocation. Simple joint capsule tear or ligament rupture is rare and often accompanied by severe cuboid compression fracture. The treatment should be focused on restoring the stability of the bony structure and the length of the medial and lateral columns. The use of a joint-spanning external fixator helps maintain reduction and restore the length of the medial and lateral columns.

High-Level Extraction of Recyclable Nanocatalysts by Using Polyphosphazene Microparticles.

Improper disposal of metal nanoparticles has caused serious environmental and pathological problems because of their active nanotoxicity. Therefore, there is an urgent need to develop a strategy for efficiently removing redundant metal nanoparticles from water, while also permitting restoration of their catalytic activities to those of pristine particles for reapplication. Herein, we present intrinsically nitrogen-rich cross-linked polyphosphazene microparticles to capture silver nanoparticles (AgNPs) from aqueous media by a simple one-step method. The described microparticles exhibit an outstanding adsorption capacity for AgNPs of approximately 59.35 mg/g, exceeding those of other adsorbents. The adsorption kinetics of AgNPs on these microparticles obeyed a pseudo-second-order kinetic model. More importantly, the recovered AgNPs maintained good catalytic activity in the reduction of methylene blue by sodium borohydride. Based on their simple preparation, high adsorption efficiency, and nondestructive effect on the catalytic activity of the recovered AgNPs, the described polyphosphazene microparticles display promising potential for the removal and recovery of AgNPs from water.

Durable superamphiphobic silica aerogel surfaces for the culture of 3D cellular spheroids.

The 3D multicellular spheroids with intact cell-cell junctions have major roles in biological research by virtue of their unique advantage of mimicking the cellular physiological environments. In this work, a durable superamphiphobic silica aerogel surface (SSAS) has been fabricated for the upward culture of 3D multicellular spheroids. Poly(3,4-ethylenedioxythiophene) (PEDOT) was first electrodeposited on a conductive steel mesh as a first template for porous silica coating. Soot particles were then applied as a second template to construct a cauliflower-like silica aerogel nanostructure. After fluorination, a hierarchical structure with re-entrant curvature was finally fabricated as a durable superamphiphobic surface. This superamphiphobic surface also presented excellent antifouling towards biomacromolecules and cells, which has been demonstrated by the successful upward culture of cell spheroids. The upward culture makes the observation of cellular behavior in situ possible, holding great potential for 3D cellular evaluation in vitro.

Myeloid-derived suppressor cells inhibit T cell activation through nitrating LCK in mouse cancers.

Potent immunosuppressive mechanisms within the tumor microenvironment contribute to the resistance of aggressive human cancers to immune checkpoint blockade (ICB) therapy. One of the main mechanisms for myeloid-derived suppressor cells (MDSCs) to induce T cell tolerance is through secretion of reactive nitrogen species (RNS), which nitrates tyrosine residues in proteins involved in T cell function. However, so far very few nitrated proteins have been identified. Here, using a transgenic mouse model of prostate cancer and a syngeneic cell line model of lung cancer, we applied a nitroproteomic approach based on chemical derivation of 3-nitrotyrosine and identified that lymphocyte-specific protein tyrosine kinase (LCK), an initiating tyrosine kinase in the T cell receptor signaling cascade, is nitrated at Tyr394 by MDSCs. LCK nitration inhibits T cell activation, leading to reduced interleukin 2 (IL2) production and proliferation. In human T cells with defective endogenous LCK, wild type, but not nitrated LCK, rescues IL2 production. In the mouse model of castration-resistant prostate cancer (CRPC) by prostate-specific deletion of Pten, p53, and Smad4, CRPC is resistant to an ICB therapy composed of antiprogrammed cell death 1 (PD1) and anticytotoxic-T lymphocyte-associated protein 4 (CTLA4) antibodies. However, we showed that ICB elicits strong anti-CRPC efficacy when combined with an RNS neutralizing agent. Together, these data identify a previously unknown mechanism of T cell inactivation by MDSC-induced protein nitration and illuminate a clinical path hypothesis for combining ICB with RNS-reducing agents in the treatment of CRPC.

Double-Helical Nanostructures with Controllable Handedness in Bulk Diblock Copolymers.

Double-helical nanostructures with controllable handedness in bulk materials is of high interest in science and technology for the design and fabrication of new materials, in particular metamaterials, which mimic their natural homologues or even show superior properties. Herein, we report the fabrication of double-helical structures with controlled handedness through the self-assembly of an achiral diblock copolymer doped with d- and l-tartaric acid (TA). The helices showed clear handedness dependence on the chirality of the TA. The chiral arrangement of different achiral tectonic units, such as nanoparticles and organic molecules, was confirmed using this helical structure as a template. The double-helical structure will provide new knowledge for understanding the function of helices, and will enable the application of these systems as chiroptical materials.