Constructing a Comprehensive Nanopattern Library through Morphological Transitions of Block Copolymer Surface Micelles via Direct Solvent Immersion.

This study establishes a comprehensive library of nanopatterns achievable by a single block copolymer (BCP), ranging from spheres to complex structures like split micelles, flower-like clusters, toroids, disordered micelle arrays, and unspecified unique shapes. The ordinary nanostructures of polystyrene-b-poly(2-vinylpyridine) (PS-b-P2VP) surface micelles deposited on a SiOx surface undergo a unique morphology transformation when immersed directly in solvents. Investigating parameters such as immersion solvents, BCP molecular weight, substrate interactions, and temperature, this work reveals the influence of these parameters on the thermodynamics and kinetics governing the morphology transformation. Additionally, the practical application of BCP nanopattern templates for fabricating metal nanostructures through direct solvent immersion of surface micelles is demonstrated. This approach offers an efficient and effective method for producing diverse nanostructures, with the potential to be employed in nanolithography, catalysts, electronics, membranes, plasmonics, and photonics.

Sodium thiosulfate does not protect ovarian reserve from cisplatin-induced gonadotoxicity†.

Cisplatin, a platinum-containing alkylating agent, is used in the treatment of various tumors owing to its potent antitumor activity. However, it causes permanent and adverse effects, particularly hearing loss and depletion of ovarian reserve. Until recently, there were no clinically available protective agents to mitigate the adverse side effects of cisplatin-induced cytotoxicity. In 2022, sodium thiosulfate (STS) was approved by the Food and Drug Administration for mitigating hearing loss in children and adolescents undergoing cisplatin treatment. Consequently, our investigation aimed to determine if STS could protect ovarian reserve against cisplatin-induced gonadotoxicity. In an ex vivo culture, the cisplatin-only group exhibited a loss of primordial follicles, while post-STS administration after cisplatin exposure effectively protected primordial follicles. However, when post-STS was administrated either 6 or 4 h after cisplatin exposure, it did not confer protection against cisplatin-induced gonadotoxicity in postnatal day 7 or adolescent mouse models. Immunofluorescence assays using γH2AX and cPARP revealed that oocytes within primordial follicles exhibited DNA damage after cisplatin exposure, irrespective of post-STS administration. This underscores the rapid and heightened sensitivity of oocytes to gonadotoxicity. In addition, oocytes demonstrated an increased expression of pCHK2 rather than pERK, suggesting that the pathway leading to oocyte death differs from the pathway observed in the inner ear cell death following cisplatin exposure. These results imply that while the administration of STS after cisplatin is highly beneficial in preventing hearing loss, it does not confer a protective effect on the ovaries in mouse models.

Structural Homogeneity of Macromolecular Networks by End-to-End Click Chemistry between Discrete Tetrahedral Star Macromolecules.

Cross-linking via the end-to-end click chemistry of multiarm star polymers creates polymer networks with minimal inhomogeneities. Although it has been suggested that the mechanical and swelling properties of such networks depend on the absence of defects, the structural details of homogeneous networks created by this method have not yet been studied at the molecular level. Here, we report the synthesis of discrete tetrahedral star macromolecules (dTSMs) composed of polylactide (PLA) arms with discrete molecular weight and sequence. Polymer networks prepared by 4 × 4 cross-linking by Cu-free strain-promoted cyclooctyne-azide click chemistry (SPAAC) reaction exhibited a high degree of swelling (>40 fold by weight) in solvents without sacrificing mechanical robustness (elastic modulus >4 kPa). The structural details of the networks were investigated by network disassembly spectrometry (NDS) using MALDI-TOF mass spectrometry. By implementing a cleavable repeating unit in the discrete PLA arms of dTSM in a sequence-specific manner, the networks could be disassembled into fragments having discrete molecular weights precisely representing their connectivity in the network. This NDS analysis confirmed that end-to-end click reactions of dTSM networks resulted in the formation of a homogeneous network above the critical concentration (∼10 w/v%) of building blocks in the solution.

Metabolic characteristics of granulosa cell tumor: role of PPARγ signaling†.

Granulosa cell tumors are relatively rare, posing challenges for comprehension and therapeutic development due to limited cases and preclinical models. Metabolic reprogramming, a hallmark of cancer, manifests in granulosa cell tumors with notable lipid accumulation and increased expression of peroxisome proliferator-activated receptor gamma (PPARγ), a key lipid metabolism regulator. The roles of these features, however, remain unclear. In our previous work, we established a granulosa cell tumor model in mice by introducing a constitutively active Pik3ca mutant in oocytes, enabling the study of predictable tumor patterns from postnatal day 50. In this study, we characterized metabolic alterations during tumorigenesis (postnatal day 8 to day 50) and tumor growth (day 50 to day 65) in this model and explored the impact of PPARγ antagonism on human granulosa cell tumor proliferation. The tumor exhibited significant lipid accumulation, with PPARγ and the proliferation marker Ki67 co-localizing at postnatal day 65. Transcriptome analysis demonstrates that pathways for lipid metabolism and mitochondrial oxidation are promoted during tumorigenesis and tumor growth, respectively. Overlappingly upregulated genes during tumorigenesis and tumor growth are associated with lipid metabolism pathways. Correspondingly, mouse granulosa cell tumor shows overexpression of peroxisome proliferator-activated receptor gamma and DGAT2 proteins at postnatal day 65. Furthermore, GW9662 reduces the proliferation of KGN human granulosa cell tumor cells and decreases the phosphorylation of AKT and SMAD3. Our findings identify metabolic abnormalities in ooPIK3CA* granulosa cell tumor model and suggest peroxisome proliferator-activated receptor gamma as a potential driver for primary granulosa cell tumor growth.

Key takeaways for knowledge expansion of early-career scientists conducting Transdisciplinary Research in Energetics and Cancer (TREC): a report from the TREC Training Workshop 2022.

The overall goal of the annual Transdisciplinary Research in Energetics and Cancer (TREC) Training Workshop is to provide transdisciplinary training for scientists in energetics and cancer and clinical care. The 2022 Workshop included 27 early-to-mid career investigators (trainees) pursuing diverse TREC research areas in basic, clinical, and population sciences. The 2022 trainees participated in a gallery walk, an interactive qualitative program evaluation method, to summarize key takeaways related to program objectives. Writing groups were formed and collaborated on this summary of the 5 key takeaways from the TREC Workshop. The 2022 TREC Workshop provided a targeted and unique networking opportunity that facilitated meaningful collaborative work addressing research and clinical needs in energetics and cancer. This report summarizes the 2022 TREC Workshop's key takeaways and future directions for innovative transdisciplinary energetics and cancer research.

Enhanced stability, formulations, and rheological properties of nanoemulsions produced with microfludization for eco-friendly process.

HYPOTHESIS: Eco-friendly processes that are emerging around the world require mass production of low-energy, low-cost nanoemulsions. The process involving the high-concentrated nanoemulsions and diluting them with a large amount of solvent can certainly save the cost; however, not much detailed research has been conducted on the stability mechanism and rheological characteristics of high-concentrated nanoemulsions. EXPERIMENTS: In this study, we produced nanoemulsions via the microfluidization (MF) process, comparing their dispersion stability and rheological characteristics with macroemulsions across various oil and surfactant concentrations. Droplet mobility and dispersion stability depended on these concentrations, with Asakura-Osawa-type attractive depletion considering interparticle interaction's role in stability changes. We investigated nanoemulsions' long-term stability based on turbidity and droplet size changes over four weeks, proposing a stability diagram showing four different states depending on emulsification conditions. FINDINGS: We explored the microstructure of emulsions under varying mixing conditions, observing their effects on droplet mobility and rheological properties. We monitored changes in rheology, turbidity, and droplet size over 4 weeks, establishing stability diagrams for macro- and nanoemulsions. The stability diagrams revealed that the stability of emulsions are sensitively dependent on the droplet size, concentrations, surfactant cocentrations and the strcture of coexistent phases in case of macroscopic segregation are significantly different depending on the droplet sizes. We identified their respective stability mechanisms and discovered the relationship between stability and rheological properties for highly concentrated nanoemulsion.

Uncovering Tumor-Promoting Roles of Activin A in Pancreatic Ductal Adenocarcinoma.

Pancreatic ductal adenocarcinoma (PDAC) is one of the most lethal cancers with high incidence rates of metastasis and cachexia. High circulating activin A, a homodimer of inhibin ßA subunits that are encoded by INHBA gene, predicts poor survival among PDAC patients. However, it still raises the question of whether activin A suppression renders favorable PDAC outcomes. Here, the authors demonstrate that activin A is abundantly detected in tumor and stromal cells on PDAC tissue microarray and mouse PDAC sections. In orthotopic male mice, activin A suppression, which is acquired by tumor-targeted Inhba siRNA using cholesterol-modified polymeric nanoparticles, retards tumor growth/metastasis and cachexia and improves survival when compared to scramble siRNA-treated group. Histologically, activin A suppression coincides with decreased expression of proliferation marker Ki67 but increased accumulation of α-SMAhigh fibroblasts and cytotoxic T cells in the tumors. In vitro data demonstrate that activin A promotes KPC cell proliferation and induces the downregulation of α-SMA and upregulation of IL-6 in pancreatic stellate cells (PSC) in the SMAD3-dependent mechanism. Moreover, conditioned media from activin A-stimulated PSC promoted KPC cell growth. Collectively, our data provide a mechanistic basis for tumor-promoting roles of activin A and support therapeutic potentials of tumor activin A suppression for PDAC.

HPV vaccine narratives on Twitter during the COVID-19 pandemic: a social network, thematic, and sentiment analysis.

INTRODUCTION: The COVID-19 pandemic has increased online interactions and the spread of misinformation. Some researchers anticipate benefits stemming from improved public awareness of the value of vaccines while others worry concerns around vaccine development and public health mandates may have damaged public trust. There is a need to understand whether the COVID-19 pandemic, vaccine development, and vaccine mandates have influenced HPV vaccine attitudes and sentiments to inform health communication strategies. METHODS: We collected 596,987 global English-language tweets from January 2019-May 2021 using Twitter's Academic Research Product track. We determined vaccine confident and hesitant networks discussing HPV immunization using social network analysis. Then, we used a neural network approach to natural language processing to measure narratives and sentiment pertaining to HPV immunization. RESULTS: Most of the tweets in the vaccine hesitant network were negative in tone (54.9%) and focused on safety concerns surrounding the HPV vaccine while most of the tweets in the vaccine confident network were neutral (51.6%) and emphasized the health benefits of vaccination. Growth in negative sentiment among the vaccine hesitant network corresponded with legislative efforts in the State of New York to mandate HPV vaccination for public school students in 2019 and the WHO declaration of COVID-19 as a Global Health Emergency in 2020. In the vaccine confident network, the number of tweets concerning the HPV vaccine decreased during the COVID-19 pandemic but in both vaccine hesitant and confident networks, the sentiments, and themes of tweets about HPV vaccine were unchanged. CONCLUSIONS: Although we did not observe a difference in narratives or sentiments surrounding the HPV vaccine during the COVID-19 pandemic, we observed a decreased focus on the HPV vaccine among vaccine confident groups. As routine vaccine catch-up programs restart, there is a need to invest in health communication online to raise awareness about the benefits and safety of the HPV vaccine.

Thyroid hormone triiodothyronine does not protect ovarian reserve from DNA damage induced by X-ray and cisplatin.

PURPOSE: Cancer therapy can induce premature ovarian insufficiency, necessitating methods for preserving fertility in female cancer patients. However, the only accepted clinical practice for doing so is cryopreservation of embryos, unfertilized ova, and ovarian tissue, despite potential options such as in vitro maturation of follicles. Therefore, considerable interest has arisen in fertoprotective agents, with research on rat ovarian granulosa cells suggesting that triiodothyronine (T3) regulates an anti-apoptosis mechanism that protects the ovarian reserve from paclitaxel-induced DNA damage. In this study, we used postnatal day 5 mouse ovary to confirm the existence of T3 thyroid hormone receptor (THR), as well as to investigate the potential protective effects of T3 against cisplatin- and X-ray-induced apoptosis. We also tested the potential anti-apoptotic effect of T3 in the breast cancer cell line MDA-MB-231. METHODS: We treated cultured mouse ovaries with varying concentration of T3 and 4 µM cisplatin and 0.2 Gy X-ray. Real-time PCR, histological analysis, immunoblot analysis, and immunofluorescence were performed to assess the potential anti-apoptotic effects of T3. RESULTS: We confirmed that THR alpha and beta are expressed in the mouse ovary. T3 (0.1, 1, 10, 100 nM, and 1 µM) does not protect ovarian reserve from cisplatin- or X-ray-induced apoptosis or DNA damage. Similarly, it does not protect mouse granulosa cells and MDA-MB-231 cells from cisplatin- or X-ray-induced apoptosis. CONCLUSION: Our findings suggest that T3 is ineffective as a fertoprotective agent, and its candidacy as a potential agent to preserve fertility should be reconsidered.

Intensified Nonequilibrium Effect of Polymer Nanocomposites with Decreasing Nanoparticle Size.

What are the most important and decisive parameters that determine the structure and the property of polymer nanocomposites (PNCs)? Previous studies answered that controlling the nanoparticle interface is critical, which can be achieved with a choice of a compatible nanoparticle, a proper surface modification, and a change in the polymer chain length. In addition to these parameters, the processing condition of PNCs has recently emerged as an influential parameter for controlling PNC properties, suggesting the existence of the nonequilibrium effect of PNCs. In this regard, we chose the solvent as a main change in the processing condition and investigated the initial solvent-driven nonequilibrium effect of PNCs with varied nanoparticle (NP) sizes. We found that the type of the initial solvent is indeed crucial in determining the ultimate properties of the PNCs, and this becomes more influential as the size of NPs decreases. The decreasing size of NPs causes a conformational change in the adsorbed polymers from tightly packed layers to loosely dangling chains. This results in much greater differences in NP microstructures and rheological properties of PNCs, indicating a stronger nonequilibrium effect with smaller NPs.

Expression and T cell regulatory action of the PD-1 immune checkpoint in the ovary and fallopian tube.

PROBLEM: Immune cell trafficking and surveillance within the ovary and fallopian tube are thought to impact fertility and also tumorigenesis in those organs. However, little is known of how native cells of the ovary and fallopian tube interact with resident immune cells. Interaction of the Programmed Cell Death Protein-1 (PD-1/PDCD-1/CD279) checkpoint with PD-L1 is associated with downregulated immune response. We have begun to address the question of whether PD-1 ligand or its receptors (PD-L1/-L2) can regulate immune cell function in these tissues of the female reproductive tract. METHOD OF STUDY: PD-1 and ligand protein expression was evaluated in human ovary and fallopian tube specimens, the latter of which included stages of tubal cell transformation and early tumorigenesis. Ovarian expression analysis included the determination of the proteins in human follicular fluid (HFF) specimens collected during in vitro fertilization procedures. Finally, checkpoint bioactivity of HFF was determined by treatment of separately-isolated human T cells and the measurement of interferon gamma (IFNγ). RESULTS: We show that membrane bound and soluble variants of PD-1 and ligands are expressed by permanent constituent cell types of the human ovary and fallopian tube, including granulosa cells and oocytes. PD-1 and soluble ligands were present in HFF at bioactive levels that control T cell PD-1 activation and IFNγ production; full-length checkpoint proteins were found to be highly enriched in HFF exosome fractions. CONCLUSION: The detection of PD-1 checkpoint proteins in the human ovary and fallopian tube suggests that the pathway is involved in immunomodulation during folliculogenesis, the window of ovulation, and subsequent egg and embryo immune-privilege. Immunomodulatory action of receptor and ligands in HFF exosomes is suggestive of an acute checkpoint role during ovulation. This is the first study in the role of PD-1 checkpoint proteins in human tubo-ovarian specimens and the first examination of its potential regulatory action in the contexts of normal and assisted reproduction.

TAp63 determines the fate of oocytes against DNA damage.

Cyclophosphamide and doxorubicin lead to premature ovarian insufficiency as an off-target effect. However, their oocyte death pathway has been debated. Here, we clarified the precise mechanism of ovarian depletion induced by cyclophosphamide and doxorubicin. Dormant oocytes instead of activated oocytes with high PI3K activity were more sensitive to cyclophosphamide. Checkpoint kinase 2 (CHK2) inhibitor rather than GNF2 protected oocytes from cyclophosphamide and doxorubicin, as cyclophosphamide up-regulated p-CHK2 and depleted primordial follicles in Abl1 knockout mice. Contrary to previous reports, TAp63 is pivotal in cyclophosphamide and doxorubicin-induced oocyte death. Oocyte-specific Trp63 knockout mice prevented primordial follicle loss and maintained reproductive function from cyclophosphamide and doxorubicin, indicated by undetectable levels of BAX and cPARP. Here, we demonstrated that TAp63 is fundamental in determining the signaling of oocyte death against DNA damage. This study establishes the role of TAp63 as a target molecule of adjuvant therapies to protect the ovarian reserve from different classes of chemotherapy.

Adipose Tissue Wasting as a Determinant of Pancreatic Cancer-Related Cachexia.

Pancreatic cancer (PC) is the third leading cause of cancer-related death in the US, and its 5-year survival rate is approximately 10%. The low survival rates largely stem from diagnostic delay and the presence of significant adipose tissue and muscle wasting, commonly referred to as cachexia. Cachexia is present in nearly 80% of PC patients and is a key cause of poor response to treatment and about 20% of death in PC patients. However, there are few clinical interventions proven to be effective against PC-related cachexia. Different cancer types feature distinct secretome profiles and functional characteristics which would lead to cachexia development differently. Therefore, here we discuss affected tissues and potential mechanisms leading to cachexia in PC. We postulate that the most affected tissue during the development of PC-related cachexia is adipose tissue, historically and still thought to be just an inert repository for excess energy in relation to cancer-related cachexia. Adipose tissue loss is considerably greater than muscle loss in quantity and shows a correlation with poor survival in PC patients. Moreover, we suggest that PC mediates adipose atrophy by accelerating adipocyte lipid turnover and fibroblast infiltration.

Oocyte CTR1 is not essential for cisplatin-induced oocyte death of primordial follicle.

Accumulated evidence indicates that cisplatin, a platinum-based alkylating agent, causes preferential DNA damage to oocytes of primordial follicles (PFs) in the ovary, suggesting oocyte-favored accumulation of cisplatin. Copper transporter 1 (CTR1; Slc31a1 ) is implicated in facilitating cisplatin uptake in cells. Here we found that oocytes of PFs had constitutively higher expression of CTR1 than other cell types in mouse ovary. However, oocyte-specific Slc31a1 knockout was not sufficient to prevent cisplatin-induced depletion of PFs in vitro . Our data indicate that CTR1 would not be the only route for cisplatin to be transported inside the oocytes of PFs in the ovary.

Tuning Surface Plasmon Resonance Responses through Size and Crosslinking Control of Multivalent Protein Binding-Capable Nanoscale Hydrogels.

Surface plasmon resonance (SPR) phenomena have been widely studied to detect biomolecules because of their high sensitivity and ability to determine biomolecular interactions with kinetic information. However, highly selective detection in specific concentration ranges relevant to target biomolecules is still a challenging task. Recently, we developed bioresponsive nanoscale hydrogels to selectively intensify SPR signals through multivalent protein binding (MPB) events with target biomolecules, including IL-2, where we were able to demonstrate exceptional selectivity for target biomolecules with minimal responses to nonspecific and monovalent binding events. In this work, we systematically explored the relationship between the physical properties of MPB-capable nanoscale hydrogels and their SPR response induced in the presence of the programmed cell death protein 1 antibody (PD-1Ab) as a model target biomolecule. First, we developed a synthetic protocol by controlling various reaction parameters to construct a library of nanoscale poly(N-isopropylacrylamide-co-acrylic acid) hydrogels (NHs) with different sizes (from 400 nm to 1 µm) and degrees of crosslinking (from 2 to 8%). Then, by incorporating MPB-capable PD-1 receptors onto the surface of NHs to form PD-1-responsive nanoscale hydrogels (PNHs), the hydrogel size and crosslinking dependency of their SPR responses were investigated. Our results reveal the appropriate hydrogel size regime and degree of crosslinking for effective PD-1Ab detection at specific concentrations range between a few nM and 1 µM. Overall, our study demonstrates that by tuning the physical properties of the nanoscale hydrogel matrix, the sensitivity and detection range of MPB-based SPR sensors can be modulated to potentially benefit clinical applications such as monitoring diverse therapeutic biomolecules.

Scaling Laws for Block Copolymer Surface Micelles.

Polymers under confinement exhibit different structures and properties from the bulk. While block copolymers (BCPs) create well-defined micelles in solution, two-dimensional (2D) spatial confinement at the air-water interface constrains the chain conformations and deforms the micellar structure, thus forming a surface micelle. The BCP surface micelles open up an opportunity in nanoscience and engineering by serving as an interfacial modifier and structural platform. Nevertheless, a scaling law, a principle governing the micellar structure, is absent. Herein, we report a unified scaling relation to describe the combinational structure of BCP surface micelles in two and three dimensions and further reveal their formation mechanism in line with the suggested scaling relation. We investigated the intrinsic scaling relations in a surface pressure-free environment by introducing a concept of excluded volume-dependent scaling exponent based on the scaling theory of 2D polymers. In addition, an extrinsic scaling relation is derived for the surface pressure-dependent corona scaling.

3D bioprinted white adipose model forin vitrostudy of cancer-associated cachexia induced adipose tissue remodeling.

Cancer-associated cachexia (CAC) is a complex metabolic and behavioral syndrome with multiple manifestations that involve systemic inflammation, weight loss, and adipose lipolysis. It impacts the quality of life of patients and is the direct cause of death in 20%-30% of cancer patients. The severity of fat loss and adipose tissue remodeling negatively correlate with patients' survival outcomes. To address the mechanism of fat loss and design potential approaches to prevent the process, it will be essential to understand CAC pathophysiology through white adipose tissue models. In the present study, an engineered human white adipose tissue (eWAT) model based on three-dimensional (3D) bioprinting was developed and induced with pancreatic cancer cell-conditioned medium (CM) to mimic the status of CACin vitro. We found that the CM induction significantly increased the lipolysis and accumulation of the extracellular matrix (ECM). The 3D eWATs were further vascularized to study the influence of vascularization on lipolysis and CAC progression, which was largely unknown. Results demonstrated that CM induction improved the angiogenesis of vascularized eWATs (veWATs), and veWATs demonstrated decreased glycerol release but increasedUCP1expression, compared to eWATs. Many unique inflammatory cytokines (IL-8, CXCL-1, GM-CSF, etc) from the CM were detected and supposed to contribute to eWAT lipolysis,UCP1up-regulation, and ECM development. In response to CM induction, eWATs also secreted inflammatory adipokines related to the metastatic ability of cancer, muscle atrophy, and vascularization (NGAL, CD54, IGFBP-2, etc). Our work demonstrated that the eWAT is a robust model for studying cachectic fat loss and the accompanying remodeling of adipose tissue. It is therefore a useful tool for future research exploring CAC physiologies and developing potential therapies.

Evidence of cancer therapy-induced chronic inflammation in the ovary across multiple species: A potential cause of persistent tissue damage and follicle depletion.

Chemotherapy and radiation treatments are known for deleterious effects on the ovary, which can result in prolonged recovery time before ovarian function resumes, including follicular growth after completion of these therapies. To better understand the protracted ovarian dysfunctions after chemotherapy and radiotherapy, we designed a comprehensive study to investigate the underlying mechanisms involved in chronic ovarian damage that prevent follicular development and/or to induce persistent follicle loss. Blood and ovarian samples were collected from reproductive age women, rhesus macaques, and mice after completion of chemotherapy and/or radiotherapy and from age-matched patients and animals without chemotherapy agent or radiation exposure to serve as controls. Serum levels of anti-Müllerian hormone and proinflammatory cytokines, monocyte chemoattractant protein 1 and IL6, were measured. Ovarian tissue was assessed for histopathology and inflammatory cell infiltration, e.g., macrophages and neutrophils, by immuohistochemistry. Serum anti-Müllerian hormone concentrations were lower, whereas proinflammatory cytokine concentrations were higher, in patients and rhesus macaques at ~1 year post-chemotherapy agent and/or radiation exposure compared with controls. The number of primordial follicles reduced in the mouse ovary > 5 weeks after a single injection of cyclophosphamide. Macrophage infiltration was observed in the ovarian cortex of humans and animals. These data suggest that chronic inflammation induced by chemotherapy agents and/or radiation treatment may be associated with persistent ovarian tissue damage, follicle depletion, and functional decline. Interventions that dampen the overactivated inflammatory response may further protect the ovary after completion of chemotherapy and radiotherapy to maintain follicle viability and support continued follicular development in female patients.

A Single Crystal 2D Hexagonal Array in a Centimeter Scale with a Self-Directed Assembly of Diblock Copolymer Spheres.

The creation of a single-grain two-dimensional (2D) nanoarray over a large area (∼1 cm2) has been only realized with expensive lithographic fabrication involving a complicated multichemical process. In this work, we report the production of a highly aligned single-grain 2D crystalline nanoarray over a centimeter-scale large area with a concept of self-directed assembly (SDA) in block copolymer (BCP) thin films. No lithographic guiding pattern is employed in SDA. A sphere-forming BCP is first transformed to transient-cylinders and aligned with shear. The aligned cylinders act as a guiding pattern to restore the sphere-morphology producing a single-grain 2D crystalline array with the following solvent vapor annealing. The SDA process has two governing parameters: orientational order of guiding patterns in the first step and the lattice matching between the transient guiding cylinders and the restored spheres. The successful application of SDA yields a single-grain of 2D crystalline hexagonal nanoarray with an exceptional long-range order, which is confirmed by employing image treating algorithms and grazing incidence small-angle X-ray scattering (GISAXS) measurements. The suggested SDA strategy is found to be effective for large-scale nanopatterning with no lithographic tools.