tbuckley/cabot-search huggingface dataset:
Transcriptomic profiling of murine GnRH neurons reveals developmental trajectories linked to human reproduction and infertility
Authors: Yassine Zouaghi, Daniel Alpern, Vincent Gardeux, Julie Russeil, Bart Deplancke, Federico Santoni, Nelly Pitteloud, Andrea Messina
Journal: Theranostics (2025)

Rationale: Neurons producing Gonadotropin-Releasing Hormone (GnRH) are essential for human reproduction and have to migrate from nose to brain during prenatal life. Impaired GnRH neuron biology results in alterations of the reproductive axis, including delayed puberty and infertility, with considerable effects on quality of life and metabolic health. Although various genes have been implicated, the molecular causes of these conditions remain elusive, with most patients lacking a genetic diagnosis. Methods: GnRH neurons and non-GnRH cells were FACS-isolated from mouse embryo microdissections to perform high-resolution transcriptomic profiling during mouse embryonic development. We analyzed our dataset to reveal GnRH neuron molecular identity, gene expression dynamics, and cell-to-cell communication. The spatial context of candidate genes was validated using in situ hybridization and spatial transcriptomic analysis. The possible links with human reproduction in health and disease were explored using enrichment analysis on GWAS data and analyzing the genetic burden of patients with congenital GnRH deficiency. Results: GnRH neurons undergo a profound transcriptional shift as they migrate from the nose to the brain and display expression trajectories associating with distinct biological processes, including cell migration, neuronal projections, and synapse formation. We revealed a timely and spatially restricted modulation of signaling pathways involving known and novel molecules, including Semaphorins and Neurexins, respectively. A particular set of genes, whose expression in GnRH neurons timely rises in late developmental stages, showed a strong association with GWAS genes linked with human reproductive onset. Finally, some of the identified trajectories harbor a diagnostic potential for congenital hypogonadism. This is supported by genetic analysis in a large cohort of patients affected by congenital GnRH deficiency, revealing a high mutation burden in patients compared to healthy controls. Conclusion: We charted the landscape of gene expression dynamics underlying murine GnRH neuron embryonic development. Our study highlights new genes in GnRH neuron development and provides novel insights linking those genes with human reproduction.
A carrier-free long-acting ropivacaine formulation using methylprednisolone sodium succinate as a dual-functional adjuvant
Authors: Weiwei Wu, Yan Wang, Feng Qiu, Yingxian Dong, Jing Liu, Yujun Zhang, Deying Gong, Yi Kang, Guoyan Zhao, Congyan Liu, Yuncheng Li, Tao Zhu, Wensheng Zhang, Guowei Che
Journal: Theranostics (2025)

Rationale: Slow-releasing formulation of local anesthetics (LAs) has been a promising strategy for pursuing long-acting opioid-free analgesia. Although many formulations based on carrier materials have been successfully developed, challenges still remain in addressing not only the biocompatibility and biodegradability issues of carrier materials, but also potential local inflammation caused by LAs. Methods: In this study, we developed a slow-releasing ropivacaine formulation based on methylprednisolone sodium succinate (MP), a small-molecule drug used in clinic. Firstly, we studied the self-assembling behavior of MP and its interaction with ropivacaine hydrochloride (RH). Then we studied how MP could manipulate the crystallization of RH and how the release profile of obtained ropivacaine crystals could be controlled. Lastly, we investigated the long-acting analgesic effect and safety of different formulations in animal models. Meanwhile we also monitored the anti-inflammatory effect of MP on cell and animal levels. Results: MP could self-assemble into nanoparticles, which could adsorb RH and induce the formation of homogeneous ropivacaine microcrystals. Higher MP ratio in the system led to the formation of smaller ropivacaine microcrystals with a moderate release rate, which generated much longer and reliable analgesic effect in animal models with considerable safety. On the other hand, MP in the formulation showed substantial anti-inflammatory effect, which was also helpful to further relieve pain and alleviate local toxicity. Conclusion: Using MP as a dual-functional adjuvant, long-acting LA formulations with considerable safety could be prepared, providing a facile solution for long-term pain management in clinic.
Nanoparticle contrast-enhanced computed tomography of sporadic aortic aneurysm and dissection: Effect of nanoparticle size and contrast agent dose
Authors: Laxman Devkota, Chen Zhang, Deborah Vela, Poonam Sarkar, Prajwal Bhandari, Zbigniew Starosolski, Renuka Menon, Dianna M Milewicz, Ying H Shen, Scott A LeMaire, Ketan B Ghaghada
Journal: Theranostics (2025)

Background: Aortopathies, such as aortic aneurysm and dissection (AAD), are associated with enhanced aortic wall permeability and endothelial dysfunction. We previously demonstrated that nanoparticle contrast-enhanced computed tomography (nCECT), which detects enhanced aortic wall permeability, could enable non-invasive detection of early AAD before its progresses to fatal aortic rupture. This study investigated the effect of nanoparticle contrast agent (NPCA) size and dose on detection of aortopathy by nCECT. Methods:In vivo studies were performed in a mouse model of sporadic AAD induced by challenging animals with high fat diet (5 weeks) and angiotensin II infusion (last one week). The effects of NPCA size (80, 150, and 240 nm) and NPCA dose (300, 600, and 1200 mg I/kg) on detection of aortopathy were studied. To examine temporal changes in aortic wall NPCA signal at sites of AAD, mice underwent longitudinal CT. To investigate changes in aortic wall integrity, mice underwent follow-up nCECT at 6 months after initial challenge. Imaging findings were compared with gross and histologic examination of the aorta. Fluorescence microscopy was used to confirm presence or absence of intramural NPCA. Results: nCECT using all three sizes of NPCAs demonstrated significantly higher sensitivity (p< 0.05) for the detection of aortopathy compared to gross examination. Histologic analysis showed excellent correlation between the nCECT finding of intramural signal and the presence of aortopathy. The absolute improvements in detection rates were 16%, 20%, and 17% for 80 nm, 150 nm, 250 nm NPCA respectively. Sensitivity of nCECT for detection of aortic injury improved with increasing NPCA dose compared to gross exam (-17% at 300 mg I/kg to 14% at 1200 mg I/kg). Temporal analysis of aortic wall NPCA signal at sites of AAD demonstrated a peak in aortic wall CT attenuation at day 3-5 post-contrast followed by gradual return to baseline by day 120. Follow-up nCECT at 6 months demonstrated absence of wall signal enhancement compared to baseline, suggesting resolution of the altered aortic wall permeability and injury. Histologic analysis demonstrated remodeling and healing of the aortic wall. Conclusions: Nanoparticle contrast-enhanced CT using all three studied nanoparticle sizes demonstrated higher sensitivity than gross examination for the detection of aortopathy. A dose-dependent effect on sensitivity was observed with only high NPCA dose (1200 mg I/kg) demonstrating superior performance compared to gross examination for detecting early stages of aortic injury. Nanoparticle contrast-enhanced CT enabled in vivo interrogation of changes in aortic wall integrity.
A distinct subgroup with Schwann-like differentiation presents immune resistance and poor prognosis in plantar melanoma
Authors: Jie Tian, Lu Zhang, Le Zhuang, Pingping Lin, Shenxi Zhang, Yicen Yan, Yu Yang, Guohong Zhang, Hang Li, Binbin Lai
Journal: Theranostics (2025)

Background: Intratumor heterogeneity in plantar melanoma orchestrates transcriptional programs that contribute to resistance to target- and immuno-therapies. However, the evolution and spatial distribution of cellular subgroups, as well as their effects on immune environment and patient prognosis, remain unclear. Methods: We analyzed 218,021 cells from 20 plantar melanoma and 6 normal samples using single-cell RNA sequencing to reveal the evolutionary characteristics and communication patterns of tumor subgroups. Spatial transcriptomics and multiplex immunohistochemistry (mIHC) were used to map the spatial distribution of these subgroups, with mIHC scores further evaluating their correlation with patient prognosis. Single-cell multiomics analysis identified key transcription factors associated with chromatin accessibility. In addition, survival analysis was performed using bulk RNA sequencing data from 68 melanoma patients. Results: We identified a continuum of subgroups originating from stem cells via transitional and Schwann cell-like precursor states, ultimately reaching a Schwann cell-like state. This evolution trajectory was supported by integrative evidence, including assessments of stemness, transitional states, RNA velocity, and transcription factors. The histological distribution of these subgroups was validated by spatial transcriptomics and multiple IHC. Notably, Schwann cell-like subgroup, regulated by transcription factor HMGA2, was associated with immune cell dysregulation and a worse prognosis, including increased invasion and lymph node metastasis. Mechanically, inhibition of HMGA2 expression blocked the transition to Schwann-like melanoma fate. Conclusions: This study reveals the unique evolutionary trajectory of plantar melanoma, showing its differentiation towards a Schwann-like fate regulated by HMGA2, leading to a decline in pigment function, enhanced immune tolerance and an increased propensity for lymph node metastasis.
An anti-FAP-scFv-functionalized exosome-carrying hydrogel delivers <i>SKI</i> mRNA to fibrotic nucleus pulposus cells to alleviate intervertebral disc degeneration by regulating FOXO3
Authors: Yueyang Li, Yu Zhai, Tianling Wang, Siya Wang, Yu Tian, Zhiqun Bian, Yang Zhang, Chao Zhang, Minghan Liu, Changqing Li
Journal: Theranostics (2025)

Rationale: Nucleus pulposus (NP) fibrosis is a contributing factor to intervertebral disc degeneration (IVDD), which lacks an effective treatment. This study focuses on elucidating the role and mechanisms of the TGF-β signaling repressor SKI in NP fibrosis and exploring its therapeutic potential. Methods: Single-cell RNA sequencing (scRNA-seq) was used to investigate fibrotic nucleus pulposus cell (NPC) subsets and assess TGF-β signaling activation. Two recombinant plasmids encoding single-chain variable fragments (scFvs) targeting the fibrotic NPC marker FAP and SKI mRNA were co-transfected into HEK-293T cells to yield functionalized exosomes (EXski+scFv). The addition of EXski+scFv to a gelatin/oxidized sodium alginate hydrogel produced a pH-responsive exosome/hydrogel system named Gel@EXski+scFv. The therapeutic effects and underlying mechanism of Gel@EXski+scFv were evaluated by RNA sequencing, molecular docking and coimmunoprecipitation. Results: A fibrotic NPC subset was characterized by elevated FAP and diminished SKI expression, along with activation of the TGF-β signaling pathway. SKI overexpression reduced fibrosis in TGF-β-treated NPCs. EXski+scFv successfully delivered SKI mRNA to FAP-expressing fibrotic NPCs. Gel@EXski+scFv possessed good mechanical properties, degradability, injectability, and biocompatibility. Gel@EXski+scFv effectively alleviated NP fibrosis and IVDD in rats. RNA sequencing, molecular docking and coimmunoprecipitation revealed that SKI could interact with FOXO3 to suppress the TGF-β signaling pathway. Conclusion: This study elucidates the mechanism by which SKI-mediated TGF-β signaling inhibition alleviates NP fibrosis. The development of the Gel@EXski+scFv system for the targeted delivery of SKI mRNA offers a promising therapeutic strategy to alleviate NP fibrosis and IVDD in the future.
Interleukin 35-producing B cells prolong the survival of GVHD mice by secreting exosomes with membrane-bound IL-35 and upregulating PD-1/LAG-3 checkpoint proteins
Authors: Jin Kyeong Choi, Evaristus C Mbanefo, Manoj Kumar Yadav, Sahar A Alhakeem, Vijayaraj Nagarajan, Natalia S Nunes, Christopher G Kanakry, Charles E Egwuagu
Journal: Theranostics (2025)

Background: Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is an effective treatment for aggressive hematologic malignancies. However, the risk of developing graft-versus-host disease (GVHD) is a significant barrier to allo-HSCT. GVHD is a debilitating condition with high mortality rates and current therapeutic options for GVHD are limited, with corticosteroids being the standard treatment. However, the adverse effects of steroids make prolonged use difficult, necessitating the development of safer therapies. IL-35-producing B-cells (i35-Bregs) have emerged as critical regulators of immunity during autoimmune diseases. In this study, we investigated whether i35-Bregs immunotherapy can suppress and mitigate GVHD. Methods: We administered a single dose of i35-Bregs (1.5×106) to mice undergoing allo-HSCT and monitored disease severity and survival of GVHD mice over 90 days post-transplantation. We discovered that i35-Bregs secrete exosomes containing membrane-bound IL-35 (i35-Exosomes) and investigated whether ex-vivo generated i35-exosomes can be used as stand-alone immunotherapy for GVHD. i35-Breg-induced expression of cytokines or checkpoint proteins (PD-1, LAG-3, CTLA-4) was analyzed by Flow cytometry, ELISA, and RNA-seq analysis. Characterization of membrane-bound IL-35 was by Proximity ligation assay (PLA), immunohistochemistry/Confocal microscopy and Alpha Fold-Multimer modeling. Results: A single dose of 1.5×106 i35-Breg reduced severity of GVHD and prolonged GVHD survival, with more than 70% i35-Breg-treated mice surviving beyond day-90 post-transplantation while observing 100% mortality among untreated mice by day-45. Contrary to the view that IL-35 is secreted cytokine, we show here that i35-Bregs mitigate GVHD via membrane-bound IL-35 and by secreting i35-exosomes. Furthermore, i35-Bregs or ex-vivo generated i35-exosomes induce alloreactive T-cells to upregulate checkpoint proteins associated with T-cell exhaustion and anergy, inhibiting alloreactive responses and propagating infectious-tolerance mechanisms that suppress GVHD. Importantly, i35-Bregs or i35-exosomes suppresses GVHD by increasing bystander lymphocytes coated with immunosuppressive i35-exosomes. Conclusions: This study demonstrates that i35-Bregs and i35-exosomes play a critical role in mitigating GVHD. The combination of i35-Breg and i35-exosome immunotherapy may be an effective strategy for treating GVHD and other inflammatory diseases.
A multifunctional injectable microsphere with enhanced near-infrared photo-antibacterial, ROS scavenging, and anti-inflammatory properties for periodontitis treatment
Authors: Ziliang Xiu, Yujing Zhu, Xiaofeng Li, Xiaoxi Jiang, Yunru Feng, Li He, Chunhui Li, Rui Cai, Gang Tao
Journal: Theranostics (2025)

Background: Pathogenic bacteria and activated immune cells in periodontal tissues continuously generate reactive oxygen species (ROS) and inflammatory mediators, creating an oxidative stress microenvironment that causes damage to cells and tissues. Antimicrobial strategies and ROS scavenging are crucial for treating periodontitis. Methods: This study encapsulated iron-curcumin nanoparticles (Fe-Cur NPs) within GelMA microspheres using microfluidic technology. The microsphere surface was then modified with a polydopamine (PDA) layer, and minocycline hydrochloride (MH) was attached to it to form multifunctional composite microspheres (GM@Fe-Cur/PDA/MH). Results: The GM@Fe-Cur/PDA/MH microspheres exhibited excellent adhesion, allowing them to remain in periodontal pockets for prolonged periods to maintain their functionality. Benefiting from the photothermal properties of PDA combined with MH, these microspheres effectively kill bacteria and mitigate excessive immune responses. Additionally, they regulate the M1/M2 polarization of macrophages, suppress the expression of inflammatory factors, and promote osteogenic protein expression under oxidative stress conditions in vitro. In a rat model of periodontitis, GM@Fe-Cur/PDA/MH effectively controlled inflammation progression and reduced alveolar bone loss. Further studies indicated that GM@Fe-Cur/PDA/MH may promote ROS scavenging by modulating the nuclear factor erythroid 2-related factor 2 (Nrf2) signaling pathway while inhibiting the activation of the nod-like receptor protein-3 (NLRP3) inflammasome. Conclusions: In summary, GM@Fe-Cur/PDA/MH represents an effective therapeutic approach that integrates antibacterial, antioxidant, anti-inflammatory, and bone loss mitigation properties, demonstrating significant potential for application in the treatment of periodontitis.
A versatile nanoplatform with excellent biofilm permeability and spatiotemporal ROS regulation for peri-implantitis treatment
Authors: Zeyu Han, Ying Li, Xin Zhan, Ming Sun, Yan Liang, Mujie Yuan, Yong Sun, Jie Cao, Baodong Zhao, Fan Li
Journal: Theranostics (2025)

Rationale: Dental implant restoration is essential for rehabilitating dentition defects. However, peri-implantitis (PI) seriously threatens the long-term stability of implants. Treating PI requires the complete eradication of plaque biofilm and the meticulous modulation of inflammatory responses. Antibacterial photodynamic therapy (aPDT) presents a promising potential in the antibacterial realm. Nonetheless, traditional aPDT for PI faces challenges such as inadequate biofilm penetration and distribution of photosensitizers, as well as a lack of precise bacteria targeting. Moreover, the excessive ROS generated by aPDT will aggravate the oxidative stress of peri-implant tissues, and this issue cannot be neglected. Methods: The CuTA-Por@ε-PL nanoplatforms (CPP NPs) were synthesized and characterized using dynamic light scattering, transmission electron microscopy, and dye probes in detail. The antibacterial and anti-inflammatory activities of CPP NPs were evaluated both in vitro and in vivo. Moreover, the in vivo therapeutic efficacy was successively analyzed through micro-CT, hematoxylin and eosin staining, Masson's staining, immunofluorescence staining, and colony formation units (CFU), among other techniques. Results: Porphyrin (Por), CuTA nanozyme with SOD/CAT activities, and ε-Polylysine (ε-PL) were combined to fabricate CPP NPs via a straightforward approach. The notable positive charge of CPP NPs facilitated biofilm penetration, distribution and precise bacteria targeting. Then, irradiation with a 660 nm laser triggered a ROS burst for biofilm elimination. After aPDT, CPP NPs scavenged the residual ROS and modulated host immunity by regulating macrophage polarization. As a result, CPP-treated groups demonstrated the most outstanding antibacterial and anti-inflammatory performance in the rat PI model. Conclusions: Given the pathogenesis of PI, this strategy rationally designed a multifunctional NP with antibacterial and anti-inflammatory functions via spatiotemporal ROS regulation. It provides a potentially novel approach for PI treatment, which may have a profound impact on improving the prognosis of patients with PI and advancing the field of implant dentistry.
Clinical value of [<sup>18</sup>F]AlF-Thretide PET/CT and early-time-point PET acquisition in the detection and staging of prostate cancer
Authors: Xin Cheng, Guozhu Hou, Mingshuai Wang, Shan Zheng, Wei Yao, Xuejuan Wang, Rong Zheng, Nianzeng Xing, Jingjing Zhang
Journal: Theranostics (2025)

Aim: Recent studies have demonstrated the potential of PET/CT with 18F-labeled ligands targeting prostate-specific membrane antigen (PSMA), as a promising method for prostate cancer (PCa) management. The aim of this study is to assess the clinical value of [18F]AlF-Thretide ([18F]AlF-PSMA-BCH) PET/CT and early time-point PET acquisition for detecting and staging PCa. Materials and Methods: From November 2022 to May 2023, a total of 73 PCa patients were included in our study. Along with whole-body PET/CT conducted at a median time of 76 min (range: 59-139 min) post-injection, a single-bed pelvic early PET/CT scan was performed, starting at a median time of 187 s (range: 161-453 s) post-injection. Visual analysis of the images was performed first, followed by semiquantitative analysis of maximum standardized uptake value (SUVmax) of primary PCa lesions, metastases, bladder, and surrounding tissues on both early and routine time-point PET/CT scans. Results: Among 56 non-surgical patients (either treatment-naive or after androgen deprivation therapy only) who had previously undergone conventional imaging, N staging was revised in 4 cases, and M staging in 2 cases. In 54 patients with bone scans for comparison, 111 lesions on [18F]AlF-Thretide PET/CT and 41 lesions on bone scans were identified as indicative of bone metastases. The median tumor-to-bladder (T/BL) ratios for primary lesions increased from 0.33 (range: 0.03-6.22) on routine time-point PET/CT to 4.66 (range: 0.24-645.00) on early time-point PET/CT. In 94.6% (53/56) of patients, the T/BL ratios were higher on early PET/CT scans than on routine time-point PET/CT scans. However, the SUVmax of surrounding tissues was found to be higher on early PET/CT scans compared to routine PET/CT scans (external iliac vessels: 8.02 ± 1.64 vs. 2.66 ± 0.59; inferior vesical artery branches near the prostate: 4.70 ± 1.09 vs. 2.30 ± 0.49; gluteus maximus muscle: 1.19 ± 0.31 vs. 0.80 ± 0.25). Of the 17 patients who underwent surgery prior to PET/CT, early PET/CT scans improved the detection rate of local recurrences from 2/17 to 5/17. Conclusion: [18F]AlF-Thretide PET/CT was shown to be a valuable imaging modality in the management of patients with PCa. Early PET/CT scans can improve the detection rate of local recurrences and provide additional information for lesions that are challenging to distinguish from urinary uptake on routine PET/CT scans.