Sympathetic nerve–fibroblast crosstalk drives nerve injury, fibroblast activation, and matrix remodeling in pancreatic cancer
Sattler et al. report that reciprocal signaling between sympathetic nerves and cancer-associated fibroblasts promotes nerve injury programs, fibroblast activation, and matrix remodeling in pancreatic cancer. The cover image shows sympathetic ganglia (cyan) interacting with activated fibroblasts (white and cyan) in vitro. Cellular nuclei are shown in dark blue. Image credit: Ariana L. Sattler.
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Research Letters
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Abstract
Authors
Johanna Tennigkeit, Maurice Wiegelmann, Chiara Massa, Jonas Lübcke, Werner Dammermann, Karina Börner, Filip Schröter, Barbara Seliger, Maximilian Kleinert, Oliver Ritter, Gregor Sachse
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Authors
Sebastian Kämpf, Marjan Hematianlarki, Leon Altmann, Jessica M. Bright, Alyssa M.A. Toda, Zohreh Mirzapoor, Valentin Zollner, Anja Werner, Johanna Bulang, Barbara Radovani, Miriam Wöhner, William Avery, Mark J. Karbarz, Pamela B. Conley, Greg P. Coffey, Falk Nimmerjahn
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Authors
Xiangmei Hua, Pranali N. Shah, Gustavo A. Velasquez, Lillian Sidky, George A. Romar, Lydia W. Boer, Natalie Hickerson, Tracy Qiying Cui, Federico Repetto, Abigail Waldman, Marilyn G. Liang, J. Paul Marcoux, MacLean Sellars, Victor Barrera, Birgitta A.R. Schmidt, Arash Mostaghimi, Ruth K. Foreman, Christine G. Lian, Sherrie J. Divito
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Physician-Scientist Development
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Abstract
Dual-degree medical students pursue additional training to prepare for careers in research, public health, and administration, but how these experiences influence residency application behaviors and outcomes are poorly understood. We analyzed 36,298 residency applicants from the Texas Seeking Transparency in Application to Residency (TexasSTAR) database spanning 2017–2023 to compare application, interview, and match patterns among single-degree MD applicants and those with MD-PhD, MD-MPH, MD-MBA, or MD-MSc degrees. Despite differences in academic metrics, application strategies, and interview rates, match rates were similar across degree groups. MD-PhD students applied to fewer programs but had the highest interview offer–to–application rate and matched at more prestigious programs based on Doximity rankings. Beyond traditional application metrics such as board scores, research productivity, grades, and honor society membership, strategies including away rotations, geographic preferencing, and program signaling were associated with increased interview offers and match success among all applicants but were less influential for dual-degree applicants. These findings suggest dual-degree applicants require specialized advising and evaluation.
Authors
Daniel C. Brock, Deborah D. Rupert, Toni Darville, Caroline S. Jansen, Elias M. Wisdom, Cynthia Y. Tang
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Research Articles
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Abstract
Dermal fibrosis is a cardinal feature of systemic sclerosis (SSc) for which there are limited effective disease-modifying therapies. SSc is characterized by dermal fibrosis accompanied by loss of dermal white adipose tissue (DWAT), yet the mechanisms linking adipocyte depletion to fibroblast activation remain unclear. Here we identify the transcription factor SIX1 as a central regulator coupling adipogenic repression with profibrotic signaling. SIX1 expression was increased in skin biopsies from 2 independent SSc cohorts and localized to fibroblast and perivascular stromal cells. In mice, ubiquitous or adipocyte-specific deletion of Six1 preserved DWAT, reduced collagen accumulation, and selectively decreased profibrotic mediators. In cultured fibroblasts, CRISPR/Cas9-mediated Six1 loss enhanced adipogenic markers while reducing profibrotic mediators and directly suppressed PAI-1 (SERPINE1) promoter activity. Together, these data position SIX1 as a transcriptional switch that promotes adipocyte reprogramming and fibrotic progression, and they highlight SIX1 inhibition as a potential therapeutic strategy to preserve adipocyte identity and limit dermal fibrosis.
Authors
Nancy Wareing, Tingting W. Mills, Scott Collum, Minghua Wu, Lucy Revercomb, René A. Girard, Hui Liu, Alexes Daquinag, Mikhail Kolonin, Marka Lyons, Brian Skaug, Weizhen Bi, Meer A. Ali, Haniyeh Koochak, Anthony R. Flores, Yuntao Yang, W. Jim Zheng, William R. Swindell, Shervin Assassi, Harry Karmouty-Quintana
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Synthetic prostaglandin analogs, such as latanoprost, are first-line treatments to reduce intraocular pressure (IOP) in the management of glaucoma, treating millions of patients daily. Glaucoma is a leading cause of blindness, characterized by progressive optic neuropathy, with elevated IOP being the sole modifiable risk factor. Despite this importance, the underlying latanoprost mechanism of action is still not well defined, being associated with both acute and long-term activities, and a growing list of ocular side effects. Prostaglandins are eicosanoid lipid mediators. Yet, there has not been a comprehensive assessment of small lipid mediators in glaucomatous eyes. Here, we performed a lipidomic screen of aqueous humor sampled from patients with glaucoma and healthy control eyes. The resulting signature was surprisingly focused on significantly elevated levels of arachidonic acid (AA) and its derivative, the antiinflammatory and cytoprotective mediator, lipoxin A4 (LXA4), in glaucomatous eyes. Subsequent experiments revealed that this response was drug induced, due to latanoprost actions on trabecular meshwork cells, rather than a consequence of elevated IOP. We demonstrate that increased LXA4 inhibited proinflammatory cues and promoted TGF-β production in the anterior chamber. In concert, an autocrine prostaglandin circuit mediated canonical rapid IOP lowering. This work reveals parallel mechanisms underlying acute and long-term latanoprost activities during glaucoma treatment.
Authors
David J. Mathew, Shubham Maurya, Julian Ho, Izhar Livne-Bar, Darren Chan, Jenny Wanyu Zhang, Yvonne M. Buys, Marisa Sit, Graham Trope, Donna M. Peters, John G. Flanagan, Karsten Gronert, Jeremy M. Sivak
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Fanconi anemia (FA) is the most common BM failure (BMF) syndrome. FA genes have a role in suppressing DNA-RNA hybrids, termed R-loops, which can be generated via transcription mediated by RNA polymerase (RNAP). How these processes, including a role in fate determination of hematopoietic stem cells (HSCs), are related to BMF is largely unknown. Single FA gene KO in mice does not recapitulate most phenotypes observed in patients with FA. Thus, we generated a mouse model for FA by introducing heterozygous Setd2, which restricts RNAP-dependent transcription. We showed that FA patient–derived cells and Setd2+/– Fanca–/– HSCs share increased R-loop and dsRNA levels and a ribosomal biogenesis defect. Further, Setd2+/– Fanca–/– HSCs displayed cell cycle arrest, mitotic errors, and BMF phenotypes. Importantly, utilizing our Setd2+/– Fanca–/– mice, we discovered that Juglone, a pan-RNAP inhibitor, reduces R-loop and dsRNA and reverses ribosomal biogenesis defects and mitotic errors, thereby rescuing BMF. This study establishes a mouse model that underscores a key role for R-loop formation, ribosomal biogenesis defects, and mitotic errors in HSCs in driving BMF in FA. We also introduce a potential therapeutic avenue based upon pan-inhibition of RNAPs utilizing Juglone.
Authors
Michihiro Hashimoto, Xiaomin Feng, Jie Bai, Huimin Zeng, Tian Li, Jue Li, Terumasa Umemoto, Paul R. Andreassen, Gang Huang
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Pancreatic cancer is a highly innervated gastrointestinal disease in which sympathetic nerves play a critical role in modulating tumor growth and the tumor microenvironment (TME). While recent studies suggest that sympathetic nerves influence various TME components, including lymphoid and myeloid immune cells, their interactions with cancer-associated fibroblasts (CAFs) remain poorly understood. CAFs are a hallmark of pancreatic tumors and are known to upregulate axon guidance and neuroactive cues, suggesting a potential feedback loop with tumor-innervating nerves. Here, we investigated the bidirectional crosstalk between sympathetic nerves and CAFs in human and mouse pancreatic tumors. Using a chemo-genetic ablation model, we selectively eliminated pancreatic sympathetic nerves and found that denervation significantly reduced tumor size in female mice. To further dissect this interaction, we established coculture systems with immortalized pancreatic fibroblasts and primary sympathetic neuron explants, identifying key transcriptional changes driven by CAF–sympathetic nerve signaling. Our findings demonstrated that sympathetic signaling enhanced CAF activation and extracellular matrix remodeling, while activated CAFs, in turn, induced transcriptional programs in sympathetic neurons associated with nerve injury response. These results establish CAFs as central mediators of the tumor-supportive role of sympathetic nerves, offering further insights into the neural regulation of pancreatic cancer progression.
Authors
Ariana L. Sattler, Parham Diba, Kevin Hawthorne, Carl Pelz, Joe Grieco, Tetiana Korzun, Bryan Chong, M.J. Kuykendall, Rosalie C. Sears, Daniel L. Marks, Mara H. Sherman, Teresa A. Zimmers, S. Ece Eksi
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Temporomandibular joint osteoarthritis (TMJOA), a prevalent subtype of temporomandibular disorders, is characterized by progressive cartilage degradation and subchondral bone destruction. Despite advancements in understanding TMJOA pathogenesis, the molecular mechanisms underlying its progression remain unclear. In this study, elevated Slit guidance ligand 2 (SLIT2) expression was observed in TMJ tissues of unilateral anterior crossbite–induced TMJOA mice and synovial fluid from patients with TMJOA, correlating with disease severity. Furthermore, SLIT2 overexpression in transgenic mice intensified TMJOA progression, whereas heterozygous deletion of roundabout guidance receptor 1/2 (ROBO1/2) alleviated cartilage and bone damage. Mechanistically, SLIT2 promoted ROBO1-LRP6 complex formation, facilitating LRP6 phosphorylation and β-catenin nuclear translocation. This cascade upregulated matrix-degrading enzymes while downregulating cartilage structural proteins, exacerbating cartilage destruction and subchondral bone loss. These findings suggest that the SLIT2/ROBO1/LRP6 axis may represent a potential therapeutic target for TMJOA and provide mechanistic insights into disease progression.
Authors
Guan Luo, Baoyi Chen, Wenjun Chen, Huiyi Lin, Weiqi Guo, Qingbin Zhang, Jiang Li, Lijing Wang, Janak Lal Pathak, Yuhui Yang, Weijun Zhang, Xiaoyu Zhang, Beining Zheng, Ziyi Wang, Shiting Wei, Jiaxin He, Wei-Jie Zhou, Chang Liu
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This study provides a comprehensive evaluation of the cutaneous adipose tissue (CAT) transcriptome in patients with psoriasis and investigates the effects of IL-17 blockade on CAT inflammation through a randomized placebo-controlled trial using secukinumab (ObePso-S study, ClinicalTrials.gov NCT03055494). RNA sequencing analysis of CAT biopsies from 82 patients with psoriasis revealed 2132 differentially expressed transcripts compared with healthy controls. Notably, significant gene dysregulation was observed in both lesional skin (LS)-CAT and non-lesional (NL)-CAT, including activation of IL-17–driven pathways, antimicrobial peptide–related, and neutrophil degranulation signatures. Stratification by obesity demonstrated that obese psoriatic CAT exhibited a more than 2-fold higher number of differentially expressed genes than non-obese counterparts, suggesting a synergistic interaction between psoriasis and obesity in driving CAT inflammation. Treatment with secukinumab markedly improved inflammatory signatures in psoriatic CAT, with greater improvements observed in obese patients. These findings reveal a pronounced and partially IL-17–dependent inflammatory phenotype in psoriatic CAT, challenge the conventional concept of psoriasis as a solely superficial skin disease, and highlight CAT as an important contributor to systemic inflammation in psoriasis.
Authors
Naomi Shishido-Takahashi, Sandra Garcet, Inna Cueto, Hong Beom Hur, Elisa Muscianisi, Jennifer Steadman, Andrew Blauvelt, James G. Krueger
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Proper development of the umbilical cord and placental vasculature is essential for embryonic development. While the allantois is known give rise to endothelial cells (ECs) within the placenta, whether the allantois gives rise to ECs in the umbilical cord is debated. Furthermore, a lack of genetic tools to study placental vascular development independent of the embryo proper has hindered robust investigation into the primary cause of vascular defects from early studies utilizing global KOs. In this study, we delineate the contribution of the allantois to the umbilical vessels and utilize a mouse genetic tool previously developed by our lab to revisit the role of Notch signaling during placental development. We show that the allantois has mosaic contribution to the umbilical endothelium with higher contributions closer to the placenta. Allantoic deletion of Dll4 disrupts umbilical cord and placental vascular formation with secondary defects in the heart. Lastly, we identify Unc5b downstream of Notch signaling that restricts EC migration while promoting chemokine signaling for vascular smooth muscle cell (vSMC) recruitment to arteries. These findings identify a genetic tool for investigating placental vascular development and give insights into the ontogeny and mechanisms of placental vascular and umbilical cord development.
Authors
Derek C. Sung, Hana A. Ahanger, Sweta Narayan, Jesse A. Pace, Mei Chen, Jisheng Yang, Siqi Gao, T.C.S. Keller IV, Jenna Bockman, Xiaowen Chen, Erica Nguyen, Alan T. Tang, Patricia Mericko-Ishizuka, Ivan Maillard, Mark L. Kahn
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Systemic sclerosis (SSc) is a complex and heterogeneous condition characterized by progressive fibrosis in multiple organs. Recent studies implicate plasminogen activator inhibitor-1 (PAI-1) in the pathogenesis of SSc, and PAI-1 is considered as a potential target for therapy. Here, using single-cell and spatial RNA-seq analysis of skin biopsies from 18 healthy individuals and 22 SSc patients, we found elevated PAI-1 colocalizing to myofibroblasts with enriched extracellular matrix–associated biological processes. Treatment of SSc dermal fibroblasts with the small-molecule PAI-1 inhibitor MDI-2517 reduced the expression of the profibrotic markers COL1A1 and ACTA2. To investigate the therapeutic potential of MDI-2517, we evaluated its efficacy in reducing fibrosis in a preclinical model of SSc. Treatment of mice with MDI-2517 significantly reduced both skin and lung fibrosis and was superior to treatment with either pirfenidone or mycophenolate mofetil. Additionally, MDI-2517 attenuated weight loss and significantly reduced the expression of key profibrotic markers. Compared with tiplaxtinin, another PAI-1 inhibitor previously shown to be effective in a model of SSc, MDI-2517 was found to have superior efficacy at a 10-fold lower dose. These findings highlight the role of PAI-1 in the pathogenesis of SSc, and the potential of MDI-2517 for the treatment of SSc.
Authors
Enming J. Su, Pei-Suen Tsou, Mark Warnock, Natalya Subbotina, Kris Mann, Sirapa Vichaikul, Alyssa Rosek, Lisa Leung, Xianying Xing, Enze Xing, Olesya Plazyo, Rachael Bogle, Lam C. Tsoi, Cory D. Emal, Dinesh Khanna, John Varga, Thomas H. Sisson, Johann E. Gudjonsson, Daniel A. Lawrence
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Aberrant polymorphonuclear neutrophil (PMN) accumulation in tissues induces chronic vascular diseases. Endothelial cells (ECs) regulate the access of PMNs into the tissue from the blood. However, the mechanisms that prevent PMNs from being activated and accumulating in the tissue, a hallmark of acute lung injury (ALI), remain elusive. We demonstrate that conditional deletion of Erg in ECs spontaneously alters the PMN transcriptome, which is enriched with genes that induce PMN recruitment, adhesion, activation, and “do not eat me” signals due to impaired synthesis of the deubiquitinase A20. Decreased A20 levels, in turn, activated the transcription factor NF-κB and the secretion of MIP2α (human homolog of IL-8) in ECs. EC-secreted MIP2α/IL-8 engaged the CXCR2 cascade on PMNs, leading to their activation and inflammatory injury. These findings were recapitulated in the lungs and blood of PMNs from patients dying of ALI. Overexpression of the A20 gene in ECs or pharmacological inhibition of CXCR2 on PMNs in iEC-Erg–/– mice rescued EC control of PMNs and tissue homeostasis, and enhanced mouse survival after pneumonia. Thus, the EC/Erg/A20 axis regulates PMN accumulation and hyperactivation in the lungs by inhibiting EC-mediated IL-8 activation of PMN CXCR2, thereby providing a potential target for neutrophilic inflammatory vascular diseases.
Authors
Vigneshwaran Vellingiri, Vijay Avin Balaji Ragunathrao, Jagdish Chandra Joshi, Md Zahid Akhter, Mumtaz Anwar, Somenath Banerjee, Sayanti Datta, Viktor Pinneker, Steven Dudek, Yoshikazu Tsukasaki, Sandra Pinho, Dolly Mehta
×Abstract
Alveolar macrophages (AMs) catabolize lipid-rich pulmonary surfactant to support gas exchange and have antiinflammatory programming to limit tissue damage in response to minor challenges. GATA transcription factors (TFs) shape immune cell fates, and GATA2 is expressed in a lung-specific manner in macrophages. GATA2 mutations and lung macrophage downregulation of GATA2 have been associated with chronic pulmonary pathologies in humans, but the role of GATA2 in coordinating AM function is not well defined. Using mice with myeloid-specific deletion of the GATA2 DNA binding C-terminal zinc finger domain, we show that GATA2 deficiency promotes enhanced inflammatory gene expression and metabolic dysfunction in AMs in response to type 2 stimuli. Although homeostatic functions of AMs remain largely intact, GATA2 deficiency increases expression of type 2 response genes during IL-33–induced inflammation. Coincident with GATA2-dependent expression of genes in metabolic pathways, Seahorse metabolic flux analysis indicates that AM metabolism is compromised in the absence of GATA2. AM GATA2-dependent gene networks are enriched for targets of TFs previously demonstrated to interact with GATA2 in other cellular contexts, including PU.1, PPARγ, and other regulators of AM function. Our data suggest that GATA2 modulates AM metabolic and transcriptomic programming to restrain responses and maintain AM identity during inflammation.
Authors
Morgan Jackson-Strong, Satarupa Ganguly, Aaron Francis, Flavia Rago, Jitendra Kanshana, Brandon A. Michalides, Lihong Teng, Omkar S. Betsur, Sonia Kruszelnicki, Karsen E. Shoger, Aaron Kim, Kay Bajpai, Amina Suleyman, Abigail Sekyere, Mika Hara, Varsha Sriram, Alok Kumar, Greg M. Delgoffe, Niranjana Natarajan, John F. Alcorn, Alison B. Kohan, Rachel A. Gottschalk
×Vancomycin eliminates gut deoxycholic acid, restoring ER proteostasis in ILC2s and relieving colitis
Abstract
Ulcerative colitis (UC) remission is marked by gut microbiota restructuring, but how microbial metabolites influence immune-mediated tissue repair is unclear. Here, we demonstrate that oral vancomycin alleviates colitis symptoms in murine models, mirroring its clinical efficacy in inducing remission in patients with UC. Mechanistically, vancomycin’s therapeutic effect is achieved by reducing deoxycholic acid (DCA). We reveal that DCA impairs mucosal repair driven by group 2 innate lymphoid cells (ILC2s) by inducing ER stress through direct binding to thioredoxin-related transmembrane protein 2 (TMX2). This interaction disrupts TMX2’s role in protein folding, triggering unresolved unfolded protein response via hyperactivation of PERK/eIF2α signaling, which suppresses the production of pro-healing molecules by ILC2s. Pharmacological inhibition of PERK phosphorylation restores ILC2 function and accelerates colitis resolution. Our work uncovers a pathogenic microbiota/DCA/ILC2 axis that obstructs mucosal healing and positions vancomycin as a targeted strategy to eliminate DCA, thereby promoting UC remission.
Authors
Qiuheng Tian, Han Liu, Xiang Gu, Jing Shen, Xi Yuan, Mengqi Zheng, Yunjiao Zhai, Yatai Chen, Penghu Han, Yangchun Ma, Wei Xin, Hongyue Ma, Yu Li, Sihan Wang, Lei Guo, Detian Yuan, Yanbo Yu, Shiyang Li
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Heart failure (HF) persists as the primary cause of death among patients recovering from acute myocardial infarction (AMI). Protein ubiquitination has been implicated as a key modulator of HF pathogenesis, yet the role of ubiquitination in the Aldh2 rs671 mutant — the most common single-nucleotide variant in human populations — remains poorly understood. We discovered TRIM21 as a previously unrecognized E3 ubiquitin ligase for the ALDH2 rs671 mutant and elucidated its mechanistic involvement in HF progression. Using Aldh2 BM chimeric mice to model AMI, we observed that WT mice transplanted with Aldh2 rs671 donor BM developed severe myocardial fibrosis and markedly reduced cardiac systolic function 2 weeks after infarction compared with controls. This phenotype arose from defective macrophage efferocytosis caused by myeloid-specific Aldh2 rs671 mutation. Through high-resolution mass spectrometry proteomics, we identified TRIM21 as the E3 ligase targeting ALDH2. TRIM21 catalyzed K48-linked ubiquitination at ALDH2 lysine 73. Macrophage-specific Trim21 knockdown via AAV-shTrim21 reversed both the exacerbated cardiac fibrosis and systolic dysfunction by restoring macrophage efferocytosis. These findings delineate the upstream E3 ubiquitin ligase and the ubiquitination site of ALDH2, revealing a potential therapeutic target for HF.
Authors
Tianrui Han, Xin Wen, Yunyun Guo, Xiangkai Zhao, Jian Zhang, Yuguo Chen, Feng Xu
×Abstract
Short QT syndrome is a heritable arrhythmia disorder linked to sudden cardiac death. We recently identified that individuals with alternating hemiplegia of childhood (AHC), a rare neurodevelopmental disorder, can exhibit shortened corrected QT intervals and elevated risk for ventricular fibrillation. This is especially true for patients with AHC heterozygous for the recurrent ATP1A3-D801N variant, though the underlying cardiac mechanism remains unclear. We hypothesized that the D801N missense impairs Na+/K+-ATPase function, causing Ca2+ overload, shortened action potential duration (APD), and arrhythmias. Using in silico modeling and patient-derived induced pluripotent stem cell cardiomyocytes (iPSC-CMsD801N), we observed shorter APD, elevated intracellular and sarcoplasmic reticulum Ca2+ levels, and delayed afterdepolarizations (DADs) compared with WT. Additionally, increased Ca²+ influx via the Na+/Ca2+ exchanger (NCX1) during depolarization was observed in iPSC-CMsD801N. Simulations and in vitro experiments suggest that reduced ATPase function accelerated inactivation of L-type Ca2+ channels. Pharmacologic inhibition of NCX1 with ORM-10103 normalized APD and reduced DADs. These findings support a Ca2+-mediated mechanism for arrhythmogenesis in ATP1A3-D801N carriers and identify NCX1 as a potential therapeutic target.
Authors
Minu-Tshyeto K. Bidzimou, Padmapriya Muralidharan, Zhushan Zhang, Danyal Raza, Daniel Needs, Bo Sun, Robin M. Perelli, Mary E. Moya-Mendez, P.K. Rakesh Manivannan, Arsen S. Hunanyan, Abbigail Helfer, Christine Q. Simmons, Alfred L. George Jr., Donald M. Bers, Nenad Bursac, Mohamad A. Mikati, Andrew P. Landstrom
×Abstract
Angiopoietin-2 (ANGPT2) is known to destabilize vascular barriers in most peripheral organs; however, its role in the brain vasculature remains poorly understood. To investigate its physiological function within the brain vasculature, we analyzed constitutive Angpt2-knockout mice in adulthood. We showed that loss of ANGPT2 leads to region-specific vascular malformations and blood-brain barrier (BBB) dysfunction, resulting in differential permeability to 1 kDa and 70 kDa fluorescent tracers. Notably, overt vascular malformations appeared only in select brain regions that allowed leakage of both tracers. These malformations were characterized by dilated, intertwined, and sprouting endothelial cells, surrounded by reactive perivascular cells, along with high levels of astrocyte- and neuron-derived vascular endothelial growth factor A (VEGFA) and elevated expression of the vascular receptors VEGF receptor 2 (KDR) and neuropilin-1 (NRP1). Other cortical areas without obvious malformations exhibited significant leakage of the 1 kDa tracer. We also demonstrated that different cell types took up the tracers after passing the BBB. Our findings identified ANGPT2 as an important factor involved in the regulation of cerebrovascular architecture, barrier integrity, and endothelial-parenchymal interactions, and uncovered surprising differences in the leakage patterns and cellular uptake of two widely used BBB tracers.
Authors
Weihan Li, Elisa Vázquez-Liébanas, Chanaëlle Fébrissy, Florent Sauvé, Jianhao Wang, Doğan E. Sayıner, Pia Buslaps, Amanda Norrén, Michael Vanlandewijck, Liqun He, Marie Jeansson, Lars Muhl, Maarja Andaloussi Mäe
×Abstract
Supernumerary centrosomes are a hallmark of cancer. To maintain viability, cancer cells cluster these centrosomes during mitosis, enabling bipolar division similar to that of normal cells. Disruption of this centrosome clustering leads to multipolar anaphase and apoptosis (anaphase catastrophe), which selectively eliminates cancer cells harboring supernumerary centrosomes. In this context, because the motor protein KIFC1 contributes to centrosome clustering, we investigated whether targeting of this mechanism through KIFC1 inhibition could be exploited in small-cell lung cancer (SCLC), an aggressive malignancy with limited treatment options and poor prognosis. Through in silico and in vitro analyses, as well as IHC of clinical samples, we found that KIFC1 is overexpressed and that centrosome amplification occurs more frequently in SCLC compared with normal tissues and other cancer types. Pharmacological and genetic inhibition of KIFC1 disrupted the clustering of supernumerary centrosomes, triggered multipolar mitosis, and exerted antineoplastic effects in SCLC cells, with minimal effects on noncancerous cells. These findings were validated and extended in vivo using SCLC xenograft models. Finally, cotargeting KIFC1 and the centrosome duplication regulator PLK4 further enhanced growth suppression in SCLC cells. Together, these results suggest that disrupting centrosome clustering and triggering anaphase catastrophe via KIFC1 inhibition may represent a promising therapeutic strategy for SCLC.
Authors
Natsuki Nakagawa, Minemichi Toda, Akiko Kunita, Masafumi Horie, Masakatsu Tokunaga, Hiroaki Ikushima, Mirei Ka, Takahiro Iida, Manabu Shigeoka, Yukinobu Ito, Takahiro Ando, Kousuke Watanabe, Yasunori Ota, Xi Liu, Ethan Dmitrovsky, Hidenori Kage, Masanori Kawakami
×Abstract
The contribution of 9p deletion to B cell acute lymphoblastic leukemia (B-ALL) has remained elusive since its discovery more than 40 years ago. Here we show that loss of CD72 is recurrent in B-ALL cases containing PAX5 deletions, and that Cd72 haploinsufficiency drives B-ALL development in Pax5+/– mice. Mechanistically, Cd72+/–;Pax5+/– precursor B cells exhibited an inflammatory transcriptional profile characterized by a decrease in Myd88 expression, a finding that aligns with our previous studies of B-ALL development in Pax5+/– mice following exposure to immune stressors. These combined genomic analyses and functional models provide compelling evidence that co-deletion of 2 contiguous genes, Pax5 and Cd72, drives B cell leukemogenesis.
Authors
Belén Ruiz-Corzo, Ana Casado-García, Ninad Oak, Paula Somoza-Cotillas, Andrea López-Álvarez de Neyra, Jorge Martínez-Cano, Alba Pérez-Pons, Elena G. Sánchez, Oscar Blanco, Diego Alonso-López, Javier De Las Rivas, Susana Riesco, Pablo Prieto-Matos, Francisco Javier García Criado, María Begoña García Cenador, Alberto Orfao, Manuel Ramírez-Orellana, César Cobaleda, Carolina Vicente-Dueñas, Kim E. Nichols, Isidro Sánchez-García
×Abstract
Adeno-associated viruses (AAVs) have been used in gene therapy, especially for inherited retinal diseases. Despite their effectiveness in gene transduction, immune responses to the AAV capsid and transgene products have been reported, which can compromise both the efficacy and the safety of AAV-mediated therapies. The eye is regarded as an immune-privileged organ where immune activity is constitutively suppressed. Here, we highlight that immunomonitoring in an ocular gene transfer reveals variable immune responses, whatever the species (human clinical trial, nonhuman primates, mice), the site of injection, the cassette, and the dose. We further explored factors contributing to this variability, investigating the potential correlation among immune parameters in a controlled experimental setting. In a syngeneic murine model after a subretinal injection of AAV, our results highlight an interindividual variability of immune parameters, emphasizing the importance of considering inherent variability among individuals when designing personalized therapies.
Authors
Duohao Ren, Gaelle A. Chauveau, Julie Vendomele, Emilie Cabon, Audrey Pineiro, Catherine Vignal-Clermont, Hanadi Saliba, Giuseppe Ronzitti, Anne Galy, Deniz Dalkara, Juliette Pulman, Divya Ail, Sylvain Fisson
×Abstract
BACKGROUND We constructed multi-trait polygenic risk scores (PRSs) predicting chronic obstructive pulmonary disease (COPD) and exacerbations, validated their performance in diverse cohorts, and identified PRS-related proteins for potential therapeutic targeting.METHODS PRSmix+, a multi-trait PRS framework, is used to train a composite PRS (PRSmulti) in COPDGene non-Hispanic White participants (n = 6,647). Associations of PRSmulti with COPD status (GOLD 2–4 vs. GOLD 0 or ICD) and exacerbation frequency were tested in COPDGene African American (n = 2,466), ECLIPSE (n = 1,858), Mass General Brigham Biobank (n = 15,152), and All of Us (n = 118,566). Protein prediction models were applied to GWAS summary statistics from traits contributing to PRSmulti and were validated with proteomic data in COPDGene (n = 5,173) and UK Biobank (n = 5,012).RESULTS PRSmix+ selected 7 traits for PRSmulti. In multivariable models, PRSmulti was associated with COPD status (meta-analysis random effects [RE] OR 1.58 [95% CI: 1.28–1.94]) and exacerbation frequency (meta-analysis RE β 0.21 [95% CI: 0.11–0.31]), with higher effect sizes observed in smoking-enriched cohorts. PRSmulti outperformed traditional single-trait PRS in all tested cohorts. Using protein prediction models, we identified 73 proteins associated with the PRSs that were also validated with measured protein levels in COPDGene and UK Biobank. Of these proteins, 25 were linked to approved or investigational drugs. Notable targets include RAGE/sRAGE, IL1RL1, and SCARF2, all implicated in COPD pathogenesis and exacerbations.CONCLUSIONS Multi-trait PRS improves prediction of COPD and exacerbation risk. Integration with proteomic data identifies druggable protein targets, offering a promising avenue for precision medicine in COPD management.TRIAL REGISTRATION COPDGene: ClinicalTrials.gov NCT00608764; ECLIPSE: ClinicalTrials.gov NCT00292552.
Authors
Chengyue Zhang, Iain R. Konigsberg, Yixuan He, Jingzhou Zhang, Tinashe Chikowore, William B. Feldman, Xiaowei Hu, Yi Ding, Bogdan Pasaniuc, Diana Chang, Qingwen Chen, Jessica A. Lasky-Su, Julian Hecker, Martin D. Tobin, Jing Chen, Sean Kalra, Katherine A. Pratte, Hae Kyung Im, Emily S. Wan, Ani Manichaikul, Edwin K. Silverman, Russell P. Bowler, Leslie A. Lange, Victor E. Ortega, Alicia R. Martin, Michael H. Cho, Matthew R. Moll
×Abstract
Giant cell aortitis (GCA) is an inflammatory disease of the aortic wall with a characteristic giant cell pattern on pathology and can lead to life-threatening aortic aneurysm and dissection. Pathogenic GCA mechanisms underlying aortic inflammation and persistence remain elusive. Here, we demonstrate the complexity of medial layer destruction and immune cell infiltration in clinical granulomatous GCA and lymphoplasmacytic IgG4-related aortitis samples using imaging-based gene expression profiling. Single-cell spatial profiling revealed aortic wall remodeling in the GCA aortas, highlighting substantial phenotypic modulation in stromal cells, including vascular smooth muscle cells (SMCs) and fibroblasts. Specifically, we observed the expansion of stromal cells expressing Tenascin-C (TNC) mRNA and spatially refined TNC accumulation in lesion areas. We confirmed these findings histologically using diseased aortas resected from individuals with giant cell arteritis and clinically isolated aortitis. Mechanistically, our data suggest that TNC promotes a proinflammatory phenotype in primary human SMCs, elevating IL-6 levels partially through the TLR4/NF-κB pathway. IL-6 signaling propagates the proinflammatory loop by activating STAT3. Pharmacological blockade of the IL-6 receptor using tocilizumab alleviated the TNC-driven proinflammatory phenotype. We propose that TNC acts as a local catalyst of inflammatory disease persistence mainly via IL-6 signaling activation and offers a potential avenue for sustained disease remission.
Authors
Hui Shi, Ying Tang, Jing Li, Ora Gewurz-Singer, Bo Yang, Dogukan Mizrak
×Abstract
The biological mechanisms underlying long COVID in the pediatric population are poorly understood. Our study aimed to characterize the immune pathophysiology of long COVID in this population. We analyzed major immune cell compartments in PBMCs and the specific SARS-CoV-2 antibody response in 99 patients with long COVID and in 18 patients without long COVID at 3 months after acute infection. Our findings indicate that pediatric long COVID is associated with a dysregulated immune response characterized by altered innate immunity and overactivated T, B, and NK cell responses. Furthermore, young people with long COVID had an impaired humoral response to SARS-CoV-2 marked by a dysregulated B cell compartment and lower levels of anti-RBD IgG and IgA. This correlated with reduced neutralizing capacity against SARS-CoV-2. Random forest analysis identified CCR6 expression on myeloid cells as the most relevant biomarker that distinguishes individuals with long COVID from control individuals with 79% accuracy.
Authors
Jon Izquierdo-Pujol, Núria Pedreño-López, Tetyana Pidkova, Maria Nevot, Victor Urrea, Fernando Laguía, Francisco Muñoz-López, Judith Dalmau, Alba Gonzalez-Aumatell, Clara Carreras-Abad, Maria Mendez, Carlos Rodrigo, Marta Massanella, Julià Blanco, Jorge Carrillo, Benjamin Trinité, Javier Martinez-Picado, Sara Morón-López
×Abstract
At-home blood collection devices (ABCDs) can facilitate study participation for remote and rural cohorts. Previous studies used ABCDs to interrogate samples by proteomics and sequencing approaches. We wanted to address the question of whether this approach could be used to assess live immune cells with high-parameter flow cytometry to enable remote immune monitoring. We first compared blood from standard venipuncture with ABCD blood draws, followed by assessment of the impact of sample shipping on immune cell viability and phenotyping. We found that capillary blood collected with a Tasso+ device and concurrently drawn venipuncture blood samples had highly congruent immune cell composition and phenotype. Shipment of Tasso+ samples via the United States Postal Service altered the myeloid compartment, but T cell numbers, subsets, and phenotypes remained remarkably stable compared with non-shipped samples. Finally, we describe a flow cytometry analysis framework that allowed for direct sample comparison even when samples were stained and analyzed over a time period of 1.5 years. Overall, our data highlight the feasibility of using ABCDs combined with subsequent flow cytometry analysis for remote immune monitoring. Additionally, our study also identifies areas that could be improved to further promote the use of ABCDs for immune monitoring.
Authors
Andrew J. Konecny, Fang Yun Lim, Eva Domenjo-Vila, Erika Lovas, Rachel L. Blazevic, Louise E. Kimball, Michael Boeckh, Alpana Waghmare, Martin Prlic
×Abstract
Antiretroviral therapy (ART) has prolonged the life expectancy of persons living with HIV, the majority of whom are now older than 50 years. Aging people with HIV are at increased risk for cardiovascular events driven by HIV-related inflammation and hypercoagulation. Apixaban is a factor Xa inhibitor that reduces cardiovascular risks and treats stroke, deep vein thrombosis, and pulmonary embolism. We assessed apixaban’s impact on key parameters of HIV/SIV pathogenesis in SIV-infected, aged rhesus macaques (RMs) receiving ART. Inflammation, coagulation, T cell subsets, B cells, and macrophages and their immune activation status were monitored throughout the study. We found no significant differences between the apixaban-treated and control groups for virus replication or CD4+ T cell recovery in blood and tissues after ART. Apixaban did not significantly affect D-dimer, immune activation, or inflammation of SIV-infected, ART-treated RMs. Apixaban-treated RMs experienced multiple bleeding episodes, tissue hemorrhages, and myocardial infarctions, as demonstrated by pathological examination of necropsy-collected tissues. Given apixaban’s lack of effect on immune activation, CD4+ T cell restoration, and inflammation, along with increased risk of hemorrhage, factor Xa inhibition may not be an efficient or safe option to target and prevent cardiovascular events in aging people with HIV.
Authors
Cuiling Xu, Haritha Annapureddy, Lilly Carson, Vansh Khurana, Ranjit Sivanandham, Sindhuja Sivanandham, Tianyu He, Kevin D. Raehtz, Janet Kim, Christie Biber, Norma Arbujas-Silva, Mohammed Daira, Sudhapriya Kandasamy, Matthew J. Feinstein, Irini Sereti, Cristian Apetrei, Ivona Pandrea
×Abstract
Current treatment protocols for most types of cancers require chemotherapeutic agents that are associated with significant side effects, including chemotherapy-induced peripheral neuropathy (CIPN). Currently, there are no effective CIPN prevention strategies, and current treatment approaches remain limited. The enzyme purine nucleoside phosphorylase (PNPase) actively modulates both oxidative injury and cellular damage. Here, we tested the hypothesis that the signs and symptoms of CIPN are due to a chemotherapy-induced dysregulation of the purine metabolome. We assessed the effect of PNPase inhibition on paclitaxel-induced (PAC-induced) CIPN. Female adult Sprague-Dawley rats were treated with PAC and randomized to oral treatment with either the PNPase inhibitor 8-aminoguanine (8-AG) or its vehicle. Some rats were injected with shRNA against PNPase prior to PAC injections. PAC-treated rats exhibited multiple abnormalities: mechanical allodynia and changes in damaging purines, intraepidermal nerve fiber (IENF) density, and signaling cascades involved in mitochondrial disruption and axonal damage. Inhibition of PNPase improved behavioral function (mechanical allodynia), rescued the loss/damage of IENF, and normalized markers for mitochondrial dysfunction and nerve damage. These findings support the hypothesis that inhibition of PNPase prevented (and potentially reversed) CIPN through several mechanisms that included a reduction in neuronal damage and development of mechanical allodynia.
Authors
Lori A. Birder, Amanda Wolf-Johnston, Jonathan Franks, Mara L.G. Sullivan, Simon C. Watkins, Anthony J. Kanai, Vladimir B. Ritov, Edwin K. Jackson
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Abstract
Inactivating NOTCH1 mutations in head and neck squamous cell carcinoma (HNSCC) were described over a decade ago, suggesting a tumorsuppressor function—unlike its oncogenic role in other tumors. Today, much debate persists regarding a putative oncogenic role in HNSCC as well, with reports that NOTCH1 signaling drives tumor growth and a cancerstemcell (CSC) phenotype. In this work, comprehensive experiments unequivocally demonstrate that NOTCH1 is a tumor suppressor in HNSCC regardless of mutation or activation status and that it reduces CSC frequency. We developed a signature of NOTCH1 activation showing the pathway is associated with very early differentiation, an altered tumor microenvironment, and better prognosis. Clarifying whether NOTCH1 occasionally functions as an oncogenic driver in HNSCC is crucial to prognosis and personalized therapy. The results presented unify the field, reconcile conflicting data, and provide critical insights into the biological and clinical significance of NOTCH1, with broader implications in other squamous carcinomas with NOTCH1 mutations.
Authors
Chenfei Huang, Shhyam Moorthy, Qiuli Li, Kazi M. Ahmed, Kalil Saab, Defeng Deng, Jiping Wang, Xiayu Rao, Jiexin Zhang, Yuanxin Xi, Jing Wang, Zhiyi Liu, Noriaki Tanaka, David A. Wheeler, Eve Shinbrot, Rami Saade, Curtis R. Pickering, Tong-Xin Xie, Adel K. El-Naggar, Abdullah A. Osman, Kunal Rai, Patrick A. Zweidler-McKay, John V. Heymach, Lauren A. Byers, Faye M. Johnson, Vlad C. Sandulache, Jeffrey N. Myers, Pedram Yadollahi, Mitchell J. Frederick
Abstract
Systemic inflammation is now recognized as a key contributor to epilepsy pathophysiology, yet the role of innate immune cells, particularly neutrophils, remains poorly defined in epilepsy. While preclinical studies in rodent models have implicated neutrophils in seizure activity, their phenotype in human epilepsy has not been thoroughly investigated. In this study, we aimed to characterize systemic inflammatory profiles and neutrophil-associated immune signatures in the blood of patients with drug-resistant epilepsy, compared to healthy controls. We identified a systemic low-grade inflammatory profile in patients, characterized by elevated neutrophil-to-lymphocyte ratio, C-reactive protein, pro-inflammatory cytokines (IL-6, CXCL8/IL-8, TNF-α), and activated neutrophils (CXCR4+CD62Llow). Neutrophil phenotyping revealed two distinct immune profiles. Patients with longer disease duration exhibited a more immature systemic signature, characterized by immature neutrophils (CD15⁺CD10⁻), resting neutrophils (CXCR4⁺CD62L⁺), and elevated IL-6 levels. In contrast, patients with higher seizure frequency displayed a more inflammatory profile, marked by increased IL-12 and activated (CXCR4+CD62Llow) and hyperactivated (CXCR4highCD62Llow) neutrophil subsets. Moreover, elevated pre-surgical levels of inflammatory profile TNF-α, IL-6, and hyperactivated CXCR4high CD62Llow neutrophils were associated with seizure recurrence one year after surgery. This pioneering study highlights the heterogeneity of peripheral immune responses in drug-resistant epilepsy and identifies neutrophil-related signatures as promising prognostic biomarkers in this context.
Authors
Coraly Simoës Da Gama, Aurelie Hanin, Gwen Goudard, Veronique Masson, Aurore Besnard, Karim Dorgham, Guy Gorochov, Guillaume Dorothee, Valerio Frazzini, Vincent Navarro, Mélanie Morin-Brureau
Abstract
The physician-scientist career has historically progressed through individual persistence and improvisation, as physician-scientists have navigated the demands of clinical practice combined with biomedical research without a clearly structured path. While this approach has sustained the field for several decades, individual determination is increasingly insufficient in the current climate, given the growing complexity within both clinical and research training, as well as potential disruptions to research funding and health care reimbursement. The 2025 ASCI / AAIM / Burroughs Wellcome Fund Physician-Scientist Pathways Workshop convened national leaders and faculty at all career stages to assess existing structures and envision new and more deliberate approaches. Discussions highlighted the impact of NIH initiatives in supporting early careers, institutional vulnerabilities, and need for intentional investments in physician-scientist careers. Breakout sessions emphasized the importance of dedicated funding for physician-scientist pathways, mentorship, social supports, and national benchmarks for compensation and promotion for this unique career pathway. The physician-scientist career path now stands at a crossroads. Going forward, sustained investment, longer and more flexible funding mechanisms such as the R37 and R35 Maximizing Investigators’ Research Award (MIRA) programs, and transparent standards are required. Federal funding alone cannot ensure the stability of a physician-scientist’s career; therefore, new approaches and commitments from academic health centers, philanthropy, and industry will be essential to ensure the viability of this career. With coordinated, intentional strategic planning, the physician-scientist workforce can thrive and remain a driver of America’s biomedical research future.
Authors
Christopher S. Williams, Emily J. Gallagher, Daniel P. Cook, David Mankoff, Rebecca M. Baron, Christopher Pittenger, Jatin M. Vyas, Don C. Rockey, Patrick J. Hu, Ashley L. Steed, W. Kimryn Rathmell, Jeffrey R. Balser, Nancy J. Brown, John M. Carethers, Jonathan A. Epstein, Keith A. Choate, Peter J. Gruber, Tiffany C. Scharschmidt, Kyu Rhee
Abstract
Malnutrition, gut inflammation, and antibiotic-induced dysbiosis (AID) are well-recognized risk factors for poor clinical outcomes among critically ill patients. We previously showed that commercially available plant-based enteral nutrition (PBEN) preserves a commensal microbiome compared with commonly used artificial enteral nutrition (AEN). In this study, PBEN was superior to AEN in promoting recovery from antibiotic-induced dysbiosis in mice and humans. PBEN effectively mitigated anemia and leukopenia, restored naïve lymphocyte populations, and reduced bone marrow myeloid expansion. Animals randomized to PBEN also exhibited improved responses to infectious challenges following antibiotic exposure. A pilot clinical study validated these findings, demonstrating increased gut commensals, reduced pathogens, and improved leukocyte balance in critically ill children receiving PBEN compared with AEN. Together, these results suggest that PBEN offers a practical dietary approach to mitigate antibiotic-associated complications and potentially improve clinical outcomes among hospitalized patients requiring supplemental nutrition.
Authors
Mona Chatrizeh, Jianmin Tian, Matthew Rogers, Firuz Feturi, Guojun Wu, Brian Firek, Roman Nikonov, Lauren Cass, Alexandra Sheppeck, Lavnish Ojha, Ali Carroll, Matthew Henkel, Justin Azar, Rajesh K. Aneja, Brian Campfield, Dennis Simon, Michael J. Morowitz
Abstract
Heart failure with preserved ejection fraction (HFpEF) is a multifactorial disease that develops in several clinical settings. Despite its complex pathogenesis, evidence indicates a central role for fibrosis in the progression of left ventricular (LV) diastolic dysfunction (LVDD). Through exploratory research into brown adipose tissue (BAT)-derived adipokines (BATokines), we identified a secreted-type pro-fibrotic protein, procollagen C-endopeptidase enhancer-1 (PCPE-1), whose expression increased in BAT with aging. PCPE-1 promotes the cleavage of procollagens and is a critical initiator of fibrillogenesis. This molecule was increased in the plasma of aged mice. In addition to aging, dietary obesity led to an increase in PCPE-1 expression in the LV of mice. Both systemic and BAT-specific PCPE-1 depletion ameliorated LV fibrosis and LVDD in the obese HFpEF model. Our data also showed that age-associated LVDD was ameliorated in the systemic PCPE-1 knockout mouse model fed with a normal chow diet. Conversely, the overexpression of PCPE-1 expression in BAT was shown to lead to aggravation of LV fibrosis and LVDD. Mechanistically, we found reactive oxygen species (ROS)/DNA damage/c-Fos/c-Jun signaling resulted in an increased production of PCPE-1 in brown adipocytes. These results indicate PCPE-1 may represent a druggable target for aging- and obesity-related HFpEF.
Authors
Yung-Ting Hsiao, Yohko Yoshida, Hirotsugu Tsuchimochi, Jingyuan Tang, Tin May Aung, Chun-Han Chang, Agian Jeffilano Barinda, Zhihong Li, Nur Syakirah Binti Othman, Tom Yoshizaki, Yiwei Ling, Shujiro Okuda, Manabu Abe, Seiya Mizuno, Satoru Takahashi, Takayuki Inomata, Hidetaka Kioka, Yasushi Sakata, Daichi Maeda, Yuya Matsue, Takaaki Furihata, Hiroshi Iwata, James T. Pearson, Kinya Otsu, Kenneth Walsh, Akihito Ishigami, Tohru Minamino, Ippei Shimizu
