@article{RN108, author = {Abitua, P. B. and Stump, L. M. and Aksel, D. C. and Schier, A. F.}, title = {Axis formation in annual killifish: Nodal and beta-catenin regulate morphogenesis without Huluwa prepatterning}, journal = {Science}, volume = {384}, number = {6700}, pages = {1105-1110}, note = {Abitua, Philip B Stump, Laura M Aksel, Deniz C Schier, Alexander F eng 2024/06/06 Science. 2024 Jun 7;384(6700):1105-1110. doi: 10.1126/science.ado7604. Epub 2024 Jun 6.}, abstract = {Axis formation in fish and amphibians typically begins with a prepattern of maternal gene products. Annual killifish embryogenesis, however, challenges prepatterning models as blastomeres disperse and then aggregate to form the germ layers and body axes. We show that huluwa, a prepatterning factor thought to break symmetry by stabilizing beta-catenin, is truncated and inactive in Nothobranchius furzeri. Nuclear beta-catenin is not selectively stabilized on one side of the blastula but accumulates in cells forming the aggregate. Blocking beta-catenin activity or Nodal signaling disrupts aggregate formation and germ layer specification. Nodal signaling coordinates cell migration, establishing an early role for this signaling pathway. These results reveal a surprising departure from established mechanisms of axis formation: Huluwa-mediated prepatterning is dispensable, and beta-catenin and Nodal regulate morphogenesis.}, keywords = {Animals *beta Catenin/metabolism Blastula/metabolism Body Patterning Cell Movement Cell Nucleus/metabolism *Fundulidae/embryology/metabolism Germ Layers/metabolism *Morphogenesis *Nodal Protein/metabolism Signal Transduction}, ISSN = {1095-9203 (Electronic) 0036-8075 (Linking)}, DOI = {10.1126/science.ado7604}, url = {https://www.ncbi.nlm.nih.gov/pubmed/38843334}, year = {2024}, type = {Journal Article} } @article{RN110, author = {Baum, M. L. and Wilton, D. K. and Fox, R. G. and Carey, A. and Hsu, Y. H. and Hu, R. and Jantti, H. J. and Fahey, J. B. and Muthukumar, A. K. and Salla, N. and Crotty, W. and Scott-Hewitt, N. and Bien, E. and Sabatini, D. A. and Lanser, T. B. and Frouin, A. and Gergits, F. and Havik, B. and Gialeli, C. and Nacu, E. and Lage, K. and Blom, A. M. and Eggan, K. and McCarroll, S. A. and Johnson, M. B. and Stevens, B.}, title = {CSMD1 regulates brain complement activity and circuit development}, journal = {Brain Behav Immun}, volume = {119}, pages = {317-332}, note = {Baum, Matthew L Wilton, Daniel K Fox, Rachel G Carey, Alanna Hsu, Yu-Han H Hu, Ruilong Jantti, Henna J Fahey, Jaclyn B Muthukumar, Allie K Salla, Nikkita Crotty, William Scott-Hewitt, Nicole Bien, Elizabeth Sabatini, David A Lanser, Toby B Frouin, Arnaud Gergits, Frederick Havik, Bjarte Gialeli, Chrysostomi Nacu, Eugene Lage, Kasper Blom, Anna M Eggan, Kevin McCarroll, Steven A Johnson, Matthew B Stevens, Beth eng Netherlands 2024/03/30 Brain Behav Immun. 2024 Jul;119:317-332. doi: 10.1016/j.bbi.2024.03.041. Epub 2024 Mar 27.}, abstract = {Complement proteins facilitate synaptic elimination during neurodevelopmental pruning, but neural complement regulation is not well understood. CUB and Sushi Multiple Domains 1 (CSMD1) can regulate complement activity in vitro, is expressed in the brain, and is associated with increased schizophrenia risk. Beyond this, little is known about CSMD1 including whether it regulates complement activity in the brain or otherwise plays a role in neurodevelopment. We used biochemical, immunohistochemical, and proteomic techniques to examine the regional, cellular, and subcellular distribution as well as protein interactions of CSMD1 in the brain. To evaluate whether CSMD1 is involved in complement-mediated synapse elimination, we examined Csmd1-knockout mice and CSMD1-knockout human stem cell-derived neurons. We interrogated synapse and circuit development of the mouse visual thalamus, a process that involves complement pathway activity. We also quantified complement deposition on synapses in mouse visual thalamus and on cultured human neurons. Finally, we assessed uptake of synaptosomes by cultured microglia. We found that CSMD1 is present at synapses and interacts with complement proteins in the brain. Mice lacking Csmd1 displayed increased levels of complement component C3, an increased colocalization of C3 with presynaptic terminals, fewer retinogeniculate synapses, and aberrant segregation of eye-specific retinal inputs to the visual thalamus during the critical period of complement-dependent refinement of this circuit. Loss of CSMD1 in vivo enhanced synaptosome engulfment by microglia in vitro, and this effect was dependent on activity of the microglial complement receptor, CR3. Finally, human stem cell-derived neurons lacking CSMD1 were more vulnerable to complement deposition. These data suggest that CSMD1 can function as a regulator of complement-mediated synapse elimination in the brain during development.}, keywords = {Animals Humans Mice *Brain/metabolism Cells, Cultured Complement C3/metabolism Complement System Proteins/metabolism *Membrane Proteins/metabolism/genetics Mice, Inbred C57BL *Mice, Knockout Microglia/metabolism *Neurons/metabolism *Synapses/metabolism Thalamus/metabolism Complement cascade Complement control protein Neural circuits Schizophrenia Synaptic pruning}, ISSN = {1090-2139 (Electronic) 0889-1591 (Linking)}, DOI = {10.1016/j.bbi.2024.03.041}, url = {https://www.ncbi.nlm.nih.gov/pubmed/38552925}, year = {2024}, type = {Journal Article} } @article{RN122, author = {Bhatia, U. and Tadman, S. and Rocha, A. and Rudraboina, R. and Contreras-Ruiz, L. and Guinan, E. C.}, title = {Allostimulation leads to emergence of a human B cell population with increased expression of HLA class I antigen presentation-associated molecules and the immunoglobulin receptor FcRL5}, journal = {Am J Transplant}, note = {Bhatia, Urvashi Tadman, Sarah Rocha, Alyssa Rudraboina, Rakesh Contreras-Ruiz, Laura Guinan, Eva C eng 2024/07/12 Am J Transplant. 2024 Jul 9:S1600-6135(24)00387-3. doi: 10.1016/j.ajt.2024.06.014.}, abstract = {In the extensive literature characterizing lymphocyte contributions to transplant-related pathologies including allograft rejection and graft-versus-host disease, T cell-focused investigation has outpaced investigation of B cells. Most B cell-related reports describe regulatory and antibody-producing functions, with less focus on the potential role of antigen-presenting capacity. Using in vitro human mixed lymphocyte reactions (MLRs) to model allostimulation, we analyzed responder B cells using transcriptional analysis, flow cytometry, and microscopy. We observed emergence of an activated responder B cell subpopulation phenotypically similar to that described in individuals with graft-versus-host disease or allograft rejection. This population had markedly increased expression of FcRL5 (Fc receptor like 5) and molecules associated with human leukocyte antigen class I antigen presentation. Consistent with this phenotype, these cells demonstrated increased internalization of irradiated cell debris and dextran macromolecules. The proportion of this subpopulation within MLR responders also correlated with emergence of activated, cytotoxic CD8(+) T cells. B cells of similar profile were quite infrequent in unstimulated blood from healthy individuals but readily identifiable in disaggregated human splenocytes and increased in both cases upon allostimulation. Further characterization of the emergence and function of this subpopulation could potentially contribute to identification of novel biomarkers and targeted therapeutics relevant to curbing transplant-related pathology.}, keywords = {FcRL5 allostimulation/allogenicity antigen cross-presentation antigen presentation graft rejection graft-versus-host disease human B cells human CD8(+) T cells}, ISSN = {1600-6143 (Electronic) 1600-6135 (Linking)}, DOI = {10.1016/j.ajt.2024.06.014}, url = {https://www.ncbi.nlm.nih.gov/pubmed/38992496}, year = {2024}, type = {Journal Article} } @article{RN111, author = {Dou, Q. and Grant, A. K. and Coutinto de Souza, P. and Moussa, M. and Nasser, I. and Ahmed, M. and Tsai, L. L.}, title = {Characterizing Metabolic Heterogeneity of Hepatocellular Carcinoma with Hyperpolarized (13)C Pyruvate MRI and Mass Spectrometry}, journal = {Radiol Imaging Cancer}, volume = {6}, number = {2}, pages = {e230056}, note = {Dou, Qianhui Grant, Aaron K Coutinto de Souza, Patricia Moussa, Marwan Nasser, Imad Ahmed, Muneeb Tsai, Leo L eng R01 CA152330/CA/NCI NIH HHS/ R01 CA169470/CA/NCI NIH HHS/ R01 EB028824/EB/NIBIB NIH HHS/ 2024/03/01 Radiol Imaging Cancer. 2024 Mar;6(2):e230056. doi: 10.1148/rycan.230056.}, abstract = {Purpose To characterize the metabolomic profiles of two hepatocellular carcinoma (HCC) rat models, track evolution of these profiles to a stimulated tumor state, and assess their effect on lactate flux with hyperpolarized (HP) carbon 13 ((13)C) MRI. Materials and Methods Forty-three female adult Fischer rats were implanted with N1S1 or McA-RH7777 HCC tumors. In vivo lactate-to-pyruvate ratio (LPR) was measured with HP (13)C MRI at 9.4 T. Ex vivo mass spectrometry was used to measure intratumoral metabolites, and Ki67 labeling was used to quantify proliferation. Tumors were first compared with three normal liver controls. The tumors were then compared with stimulated variants via off-target hepatic thermal ablation treatment. All comparisons were made using the Mann-Whitney test. Results HP (13)C pyruvate MRI showed greater LPR in N1S1 tumors compared with normal liver (mean [SD], 0.564 +/- 0.194 vs 0.311 +/- 0.057; P < .001 [n = 9]), but not for McA-RH7777 (P = .44 [n = 8]). Mass spectrometry confirmed that the glycolysis pathway was increased in N1S1 tumors and decreased in McA-RH7777 tumors. The pentose phosphate pathway was also decreased only in McA-RH7777 tumors. Increased proliferation in stimulated N1S1 tumors corresponded to a net increase in LPR (six stimulated vs six nonstimulated, 0.269 +/- 0.148 vs 0.027 +/- 0.08; P = .009), but not in McA-RH7777 (eight stimulated vs six nonstimulated, P = .13), despite increased proliferation and metastases. Mass spectrometry demonstrated relatively increased lactate production with stimulation in N1S1 tumors only. Conclusion Two HCC subtypes showed divergent glycolytic dependency at baseline and during transformation to a high proliferation state. This metabolic heterogeneity in HCC should be considered with use of HP (13)C MRI for diagnosis and tracking. Keywords: Molecular Imaging-Probe Development, Liver, Abdomen/GI, Oncology, Hepatocellular Carcinoma (c) RSNA, 2024 See also commentary by Ohliger in this issue.}, keywords = {Rats Female Animals *Carcinoma, Hepatocellular/diagnostic imaging *Liver Neoplasms/diagnostic imaging Pyruvic Acid/metabolism Magnetic Resonance Imaging Rats, Inbred F344 Lactates Abdomen/GI Hepatocellular Carcinoma Liver Molecular Imaging-Probe Development Oncology}, ISSN = {2638-616X (Electronic) 2638-616X (Linking)}, DOI = {10.1148/rycan.230056}, url = {https://www.ncbi.nlm.nih.gov/pubmed/38426887}, year = {2024}, type = {Journal Article} } @article{RN115, author = {Fukuta, T. and Kumbhojkar, N. and Prakash, S. and Shaha, S. and Da Silva-Candal, A. and Park, K. S. and Mitragotri, S.}, title = {Immunotherapy against glioblastoma using backpack-activated neutrophils}, journal = {Bioengineering & Translational Medicine}, note = {C5d1e Times Cited:0 Cited References Count:51}, abstract = {Immune checkpoint inhibitors (ICIs) represent new therapeutic candidates against glioblastoma multiforme (GBM); however, their efficacy is clinically limited due to both local and systemic immunosuppressive environments. Hence, therapeutic approaches that stimulate local and systemic immune environments can improve the efficacy of ICIs. Here, we report an adoptive cell therapy employing neutrophils (NE) that are activated via surface attachment of drug-free disk-shaped backpacks, termed Cyto-Adhesive Micro-Patches (CAMPs) for treating GBM. CAMP-adhered neutrophils (NE/CAMPs) significantly improved the efficacy of an anti-PD1 antibody (aPD-1) in a subcutaneous murine GBM model (GL261). A combination of NE/CAMPs and aPD-1 completely regressed subcutaneous GL261 tumors in mice. The efficacy of NE/CAMPs against GBM was also tested in an orthotopic GL261 model. Neutrophil's ability to migrate into the brain was not affected by CAMP attachment, and intracerebral NE/CAMP accumulation was observed in mice-bearing orthotopic GBM. The combination treatment of NE/CAMPs and aPD-1 activated systemic immune responses mediated by T cells and showed improved therapeutic responses compared with aPD-1 alone in the orthotopic GBM model. These results suggest that immunomodulation with NE/CAMPs offers a potential approach for the treatment of GBM by combination with ICIs.}, keywords = {cell therapy glioblastoma immune checkpoint inhibitor immunotherapy neutrophils t-cell exhaustion growth}, DOI = {10.1002/btm2.10712}, url = {://WOS:001289558800001}, year = {2024}, type = {Journal Article} } @article{RN107, author = {Han, X. and Lu, X. and Li, P. H. and Wang, S. and Schalek, R. and Meirovitch, Y. and Lin, Z. and Adhinarta, J. and Murray, K. D. and MacNiven, L. M. and Berger, D. R. and Wu, Y. and Fang, T. and Meral, E. S. and Asraf, S. and Ploegh, H. and Pfister, H. and Wei, D. and Jain, V. and Trimmer, J. S. and Lichtman, J. W.}, title = {Multiplexed volumetric CLEM enabled by scFvs provides insights into the cytology of cerebellar cortex}, journal = {Nat Commun}, volume = {15}, number = {1}, pages = {6648}, note = {Han, Xiaomeng Lu, Xiaotang Li, Peter H Wang, Shuohong Schalek, Richard Meirovitch, Yaron Lin, Zudi Adhinarta, Jason Murray, Karl D MacNiven, Leah M Berger, Daniel R Wu, Yuelong Fang, Tao Meral, Elif Sevde Asraf, Shadnan Ploegh, Hidde Pfister, Hanspeter Wei, Donglai Jain, Viren Trimmer, James S Lichtman, Jeff W eng UG3 MH123386/MH/NIMH NIH HHS/ U19 NS104653/NS/NINDS NIH HHS/ K99 MH128891/MH/NIMH NIH HHS/ P50 MH094271/MH/NIMH NIH HHS/ U24 NS109113/NS/NINDS NIH HHS/ England 2024/08/06 Nat Commun. 2024 Aug 5;15(1):6648. doi: 10.1038/s41467-024-50411-z.}, abstract = {Mapping neuronal networks is a central focus in neuroscience. While volume electron microscopy (vEM) can reveal the fine structure of neuronal networks (connectomics), it does not provide molecular information to identify cell types or functions. We developed an approach that uses fluorescent single-chain variable fragments (scFvs) to perform multiplexed detergent-free immunolabeling and volumetric-correlated-light-and-electron-microscopy on the same sample. We generated eight fluorescent scFvs targeting brain markers. Six fluorescent probes were imaged in the cerebellum of a female mouse, using confocal microscopy with spectral unmixing, followed by vEM of the same sample. The results provide excellent ultrastructure superimposed with multiple fluorescence channels. Using this approach, we documented a poorly described cell type, two types of mossy fiber terminals, and the subcellular localization of one type of ion channel. Because scFvs can be derived from existing monoclonal antibodies, hundreds of such probes can be generated to enable molecular overlays for connectomic studies.}, keywords = {Animals Female Mice *Cerebellar Cortex/metabolism/cytology/ultrastructure Microscopy, Confocal/methods Microscopy, Electron/methods Connectome/methods Neurons/metabolism/ultrastructure Fluorescent Dyes/chemistry Mice, Inbred C57BL Cytology}, ISSN = {2041-1723 (Electronic) 2041-1723 (Linking)}, DOI = {10.1038/s41467-024-50411-z}, url = {https://www.ncbi.nlm.nih.gov/pubmed/39103318}, year = {2024}, type = {Journal Article} } @article{RN114, author = {Hoglund, Z. and Ruiz-Uribe, N. and Del Sastre, E. and Woost, B. and Bader, E. and Bailey, J. and Hyman, B. T. and Zwang, T. and Bennett, R. E.}, title = {Brain vasculature accumulates tau and is spatially related to tau tangle pathology in Alzheimer's disease}, journal = {Acta Neuropathol}, volume = {147}, number = {1}, pages = {101}, note = {Hoglund, Zachary Ruiz-Uribe, Nancy Del Sastre, Eric Woost, Benjamin Bader, Elizabeth Bailey, Joshua Hyman, Bradley T Zwang, Theodore Bennett, Rachel E eng R00AG068602/AG/NIA NIH HHS/ R00AG061259/AG/NIA NIH HHS/ P30AG062421/AG/NIA NIH HHS/ FPU18/00630/Ministerio de Ciencia, Innovacion y Universidades/ P30 AG062421/AG/NIA NIH HHS/ 23AARG-1029355/ALZ/Alzheimer's Association/ Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Germany 2024/06/17 Acta Neuropathol. 2024 Jun 17;147(1):101. doi: 10.1007/s00401-024-02751-9.}, abstract = {Insoluble pathogenic proteins accumulate along blood vessels in conditions of cerebral amyloid angiopathy (CAA), exerting a toxic effect on vascular cells and impacting cerebral homeostasis. In this work, we provide new evidence from three-dimensional human brain histology that tau protein, the main component of neurofibrillary tangles, can similarly accumulate along brain vascular segments. We quantitatively assessed n = 6 Alzheimer's disease (AD), and n = 6 normal aging control brains and saw that tau-positive blood vessel segments were present in all AD cases. Tau-positive vessels are enriched for tau at levels higher than the surrounding tissue and appear to affect arterioles across cortical layers (I-V). Further, vessels isolated from these AD tissues were enriched for N-terminal tau and tau phosphorylated at T181 and T217. Importantly, tau-positive vessels are associated with local areas of increased tau neurofibrillary tangles. This suggests that accumulation of tau around blood vessels may reflect a local clearance failure. In sum, these data indicate that tau, like amyloid beta, accumulates along blood vessels and may exert a significant influence on vasculature in the setting of AD.}, keywords = {Humans *Alzheimer Disease/pathology/metabolism *tau Proteins/metabolism *Neurofibrillary Tangles/pathology/metabolism *Brain/pathology/metabolism Female Male Aged Aged, 80 and over Middle Aged Phosphorylation Alzheimer's disease Blood vessels Cerebral amyloid angiopathy Cerebral vasculature Neurofibrillary tangles Tau}, ISSN = {1432-0533 (Electronic) 0001-6322 (Print) 0001-6322 (Linking)}, DOI = {10.1007/s00401-024-02751-9}, url = {https://www.ncbi.nlm.nih.gov/pubmed/38884806}, year = {2024}, type = {Journal Article} } @article{RN117, author = {Janes, M. E. and Park, K. S. and Gottlieb, A. P. and Curreri, A. and Adebowale, K. and Kim, J. and Mitragotri, S.}, title = {Dendritic Cell Immune Modulation via Polyphenol Membrane Coatings}, journal = {ACS Appl Mater Interfaces}, volume = {16}, number = {22}, pages = {28070-28079}, note = {Janes, Morgan E Park, Kyung Soo Gottlieb, Alexander P Curreri, Alexander Adebowale, Kolade Kim, Jayoung Mitragotri, Samir eng 2024/05/23 ACS Appl Mater Interfaces. 2024 Jun 5;16(22):28070-28079. doi: 10.1021/acsami.4c01575. Epub 2024 May 23.}, abstract = {Cellular hitchhiking is an emerging strategy for the in vivo control of adoptively transferred immune cells. Hitchhiking approaches are primarily mediated by adhesion of nano and microparticles to the cell membrane, which conveys an ability to modulate transferred cells via local drug delivery. Although T cell therapies employing this strategy have progressed into the clinic, phagocytic cells including dendritic cells (DCs) are much more challenging to engineer. DC vaccines hold great potential for a spectrum of diseases, and the combination drug delivery is an attractive strategy to manipulate their function and overcome in vivo plasticity. However, DCs are not compatible with current hitchhiking approaches due to their broad phagocytic capacity. In this work, we developed and validated META (membrane engineering using tannic acid) to enable DC cellular hitchhiking for the first time. META employs the polyphenol tannic acid (TA) to facilitate supramolecular assembly of protein drug cargoes on the cell membrane, enabling the creation of cell surface-bound formulations for local drug delivery to carrier DCs. We optimized META formulations to incorporate and release protein cargoes with varying physical properties alone and in combination and to preserve DC viability and critical functions such as migration. We further show that META loaded with either a pro- or anti-inflammatory cargo can influence the carrier cell phenotype, thus demonstrating the flexibility of the approach for applications from cancer to autoimmune disease. Overall, this approach illustrates a new platform for the local control of phagocytic immune cells as a next step to advance DC therapies in the clinic.}, keywords = {*Dendritic Cells/drug effects/immunology *Tannins/chemistry/pharmacology *Polyphenols/chemistry/pharmacology Humans Animals Coated Materials, Biocompatible/chemistry/pharmacology Mice Cell Membrane/metabolism Cell Survival/drug effects cell therapy dendritic cells immunotherapy phenolic network vaccines}, ISSN = {1944-8252 (Electronic) 1944-8244 (Linking)}, DOI = {10.1021/acsami.4c01575}, url = {https://www.ncbi.nlm.nih.gov/pubmed/38779939}, year = {2024}, type = {Journal Article} } @article{RN109, author = {Kapate, N. and Dunne, M. and Gottlieb, A. P. and Mukherji, M. and Suja, V. C. and Prakash, S. and Park, K. S. and Kumbhojkar, N. and Guerriero, J. L. and Mitragotri, S.}, title = {Polymer Backpack-Loaded Tissue Infiltrating Monocytes for Treating Cancer}, journal = {Adv Healthc Mater}, pages = {e2304144}, note = {Kapate, Neha Dunne, Michael Gottlieb, Alexander P Mukherji, Malini Suja, Vineeth Chandran Prakash, Supriya Park, Kyung Soo Kumbhojkar, Ninad Guerriero, Jennifer L Mitragotri, Samir eng ECCS-2025158/National Science Foundation/ 1122374/National Science Foundation Graduate Research Fellowship/ NA/Harvard School of Engineering and Applied Sciences/ Germany 2024/04/06 Adv Healthc Mater. 2024 Apr 6:e2304144. doi: 10.1002/adhm.202304144.}, abstract = {Adoptive cell therapies are dramatically altering the treatment landscape of cancer. However, treatment of solid tumors remains a major unmet need, in part due to limited adoptive cell infiltration into the tumor and in part due to the immunosuppressive tumor microenvironment. The heterogeneity of tumors and presence of nonresponders also call for development of antigen-independent therapeutic approaches. Myeloid cells offer such an opportunity, given their large presence in the immunosuppressive tumor microenvironment, such as in triple negative breast cancer. However, their therapeutic utility is hindered by their phenotypic plasticity. Here, the impressive trafficking ability of adoptively transferred monocytes is leveraged into the immunosuppressive 4T1 tumor to develop an antitumor therapy. To control monocyte differentiation in the tumor microenvironment, surface-adherent "backpacks" stably modified with interferon gamma (IFNgamma) are developed to stimulate macrophage plasticity into a pro-inflammatory, antitumor phenotype, a strategy as referred to as Ornate Polymer backpacks on Tissue Infiltrating Monocytes (OPTIMs). Treatment with OPTIMs substantially reduces tumor burden in a mouse 4T1 model and significantly increases survival. Cytokine and immune cell profiling reveal that OPTIMs remodeled the tumor microenvironment into a pro-inflammatory state.}, keywords = {cancer cell therapy immunotherapy microparticles monocytes}, ISSN = {2192-2659 (Electronic) 2192-2640 (Linking)}, DOI = {10.1002/adhm.202304144}, url = {https://www.ncbi.nlm.nih.gov/pubmed/38581301}, year = {2024}, type = {Journal Article} } @article{RN112, author = {Kumbhojkar, N. and Prakash, S. and Fukuta, T. and Adu-Berchie, K. and Kapate, N. and An, R. and Darko, S. and Chandran Suja, V. and Park, K. S. and Gottlieb, A. P. and Bibbey, M. G. and Mukherji, M. and Wang, L. L. and Mooney, D. J. and Mitragotri, S.}, title = {Neutrophils bearing adhesive polymer micropatches as a drug-free cancer immunotherapy}, journal = {Nat Biomed Eng}, volume = {8}, number = {5}, pages = {579-592}, note = {Kumbhojkar, Ninad Prakash, Supriya Fukuta, Tatsuya Adu-Berchie, Kwasi Kapate, Neha An, Rocky Darko, Solomina Chandran Suja, Vineeth Park, Kyung Soo Gottlieb, Alexander P Bibbey, Michael Griffith Mukherji, Malini Wang, Lily Li-Wen Mooney, David J Mitragotri, Samir eng School of Engineering/Harvard University | Faculty of Arts and Sciences (Harvard Faculty of Arts and Sciences)/ ECCS-2025158/National Science Foundation (NSF)/ 1122374/National Science Foundation (NSF)/ 2140743/National Science Foundation (NSF)/ England 2024/03/01 Nat Biomed Eng. 2024 May;8(5):579-592. doi: 10.1038/s41551-024-01180-z. Epub 2024 Feb 29.}, abstract = {Tumour-associated neutrophils can exert antitumour effects but can also assume a pro-tumoural phenotype in the immunosuppressive tumour microenvironment. Here we show that neutrophils can be polarized towards the antitumour phenotype by discoidal polymer micrometric 'patches' that adhere to the neutrophils' surfaces without being internalized. Intravenously administered micropatch-loaded neutrophils accumulated in the spleen and in tumour-draining lymph nodes, and activated splenic natural killer cells and T cells, increasing the accumulation of dendritic cells and natural killer cells. In mice bearing subcutaneous B16F10 tumours or orthotopic 4T1 tumours, intravenous injection of the micropatch-loaded neutrophils led to robust systemic immune responses, a reduction in tumour burden and improvements in survival rates. Micropatch-activated neutrophils combined with the checkpoint inhibitor anti-cytotoxic T-lymphocyte-associated protein 4 resulted in strong inhibition of the growth of B16F10 tumours, and in complete tumour regression in one-third of the treated mice. Micropatch-loaded neutrophils could provide a potent, scalable and drug-free approach for neutrophil-based cancer immunotherapy.}, keywords = {Animals *Neutrophils/immunology *Immunotherapy/methods Mice *Polymers/chemistry *Mice, Inbred C57BL Cell Line, Tumor Tumor Microenvironment/drug effects Female Mice, Inbred BALB C Melanoma, Experimental/immunology/therapy/pathology Neoplasms/immunology/therapy Killer Cells, Natural/immunology Humans}, ISSN = {2157-846X (Electronic) 2157-846X (Linking)}, DOI = {10.1038/s41551-024-01180-z}, url = {https://www.ncbi.nlm.nih.gov/pubmed/38424352}, year = {2024}, type = {Journal Article} } @article{RN113, author = {Maddaloni, G. and Chang, Y. J. and Senft, R. A. and Dymecki, S. M.}, title = {Adaptation to photoperiod via dynamic neurotransmitter segregation}, journal = {Nature}, volume = {632}, number = {8023}, pages = {147-156}, note = {Maddaloni, G Chang, Y J Senft, R A Dymecki, S M eng England 2024/07/18 Nature. 2024 Aug;632(8023):147-156. doi: 10.1038/s41586-024-07692-7. Epub 2024 Jul 17.}, abstract = {Changes in the amount of daylight (photoperiod) alter physiology and behaviour(1,2). Adaptive responses to seasonal photoperiods are vital to all organisms-dysregulation associates with disease, including affective disorders(3) and metabolic syndromes(4). The circadian rhythm circuitry is implicated in such responses(5,6), yet little is known about the precise cellular substrates that underlie phase synchronization to photoperiod change. Here we identify a brain circuit and system of axon branch-specific and reversible neurotransmitter deployment that are critical for behavioural and sleep adaptation to photoperiod. A type of neuron called mrEn1-Pet1(7) in the mouse brainstem median raphe nucleus segregates serotonin from VGLUT3 (also known as SLC17A8, a proxy for glutamate) to different axonal branches that innervate specific brain regions involved in circadian rhythm and sleep-wake timing(8,9). This branch-specific neurotransmitter deployment did not distinguish between daylight and dark phase; however, it reorganized with change in photoperiod. Axonal boutons, but not cell soma, changed neurochemical phenotype upon a shift away from equinox light/dark conditions, and these changes were reversed upon return to equinox conditions. When we genetically disabled Vglut3 in mrEn1-Pet1 neurons, sleep-wake periods, voluntary activity and clock gene expression did not synchronize to the new photoperiod or were delayed. Combining intersectional rabies virus tracing and projection-specific neuronal silencing, we delineated a preoptic area-to-mrEn1Pet1 connection that was responsible for decoding the photoperiodic inputs, driving the neurotransmitter reorganization and promoting behavioural synchronization. Our results reveal a brain circuit and periodic, branch-specific neurotransmitter deployment that regulates organismal adaptation to photoperiod change.}, keywords = {Animals Female Mice *Adaptation, Physiological/physiology Amino Acid Transport Systems, Acidic/deficiency/genetics/metabolism *Axons/metabolism/physiology *Circadian Rhythm/physiology CLOCK Proteins/genetics Darkness Dorsal Raphe Nucleus/cytology/metabolism Neural Pathways/physiology *Neurotransmitter Agents/metabolism *Photoperiod Preoptic Area/cytology/metabolism Presynaptic Terminals/metabolism/physiology Rabies virus Serotonin/metabolism Sleep/physiology Wakefulness/physiology}, ISSN = {1476-4687 (Electronic) 0028-0836 (Linking)}, DOI = {10.1038/s41586-024-07692-7}, url = {https://www.ncbi.nlm.nih.gov/pubmed/39020173}, year = {2024}, type = {Journal Article} } @article{RN121, author = {Mapalo, M. A. and Wolfe, J. M. and Ortega-Hernandez, J.}, title = {Cretaceous amber inclusions illuminate the evolutionary origin of tardigrades}, journal = {Commun Biol}, volume = {7}, number = {1}, pages = {953}, note = {Mapalo, Marc A Wolfe, Joanna M Ortega-Hernandez, Javier eng DEB #1856679/National Science Foundation (NSF)/ England 2024/08/07 Commun Biol. 2024 Aug 6;7(1):953. doi: 10.1038/s42003-024-06643-2.}, abstract = {Tardigrades are a diverse phylum of microscopic invertebrates widely known for their extreme survival capabilities. Molecular clocks suggest that tardigrades diverged from other panarthropods before the Cambrian, but their fossil record is extremely sparse. Only the fossil tardigrades Milnesium swolenskyi (Late Cretaceous) and Paradoryphoribius chronocaribbeus (Miocene) have resolved taxonomic positions, restricting the availability of calibration points for estimating for the origin of this phylum. Here, we revise two crown-group tardigrades from Canadian Cretaceous-aged amber using confocal fluorescence microscopy, revealing critical morphological characters that resolve their taxonomic positions. Formal morphological redescription of Beorn leggi reveals that it features Hypsibius-type claws. We also describe Aerobius dactylus gen. et sp. nov. based on its unique combination of claw characters. Phylogenetic analyses indicate that Beo. leggi and Aer. dactylus belong to the eutardigrade superfamily Hypsibioidea, adding a critical fossil calibration point to investigate tardigrade origins. Our molecular clock estimates suggest an early Paleozoic diversification of crown-group Tardigrada and highlight the importance of Beo. leggi as a calibration point that directly impacts estimates of shallow nodes. Our results suggest that independent terrestrialization of eutardigrades and heterotardigrades occurred around the end-Carboniferous and Lower Jurassic, respectively. These estimates also provide minimum ages for convergent acquisition of cryptobiosis.}, keywords = {Animals *Amber *Fossils/anatomy & histology *Tardigrada/classification/anatomy & histology/genetics *Biological Evolution *Phylogeny Canada}, ISSN = {2399-3642 (Electronic) 2399-3642 (Linking)}, DOI = {10.1038/s42003-024-06643-2}, url = {https://www.ncbi.nlm.nih.gov/pubmed/39107512}, year = {2024}, type = {Journal Article} } @article{RN123, author = {Noonan, H. R. and Thornock, A. M. and Barbano, J. and Xifaras, M. E. and Baron, C. S. and Yang, S. and Koczirka, K. and McConnell, A. M. and Zon, L. I.}, title = {A chronic signaling TGFb zebrafish reporter identifies immune response in melanoma}, journal = {Elife}, volume = {13}, note = {Noonan, Haley R Thornock, Alexandra M Barbano, Julia Xifaras, Michael E Baron, Chloe S Yang, Song Koczirka, Katherine McConnell, Alicia M Zon, Leonard I eng NCI R01CA103846/NH/NIH HHS/ P01CA163222/NH/NIH HHS/ P01 CA163222/CA/NCI NIH HHS/ R01 CA103846/CA/NCI NIH HHS/ Grant given via University of Edinburgh United Kingdom project number TBD/MRA/Melanoma Research Alliance/ England 2024/06/14 Elife. 2024 Jun 14;13:e83527. doi: 10.7554/eLife.83527.}, abstract = {Developmental signaling pathways associated with growth factors such as TGFb are commonly dysregulated in melanoma. Here we identified a human TGFb enhancer specifically activated in melanoma cells treated with TGFB1 ligand. We generated stable transgenic zebrafish with this TGFb Induced Enhancer driving green fluorescent protein (TIE:EGFP). TIE:EGFP was not expressed in normal melanocytes or early melanomas but was expressed in spatially distinct regions of advanced melanomas. Single-cell RNA-sequencing revealed that TIE:EGFP(+) melanoma cells down-regulated interferon response while up-regulating a novel set of chronic TGFb target genes. ChIP-sequencing demonstrated that AP-1 factor binding is required for activation of chronic TGFb response. Overexpression of SATB2, a chromatin remodeler associated with tumor spreading, showed activation of TGFb signaling in early melanomas. Confocal imaging and flow cytometric analysis showed that macrophages localize to TIE:EGFP(+) regions and preferentially phagocytose TIE:EGFP(+) melanoma cells compared to TIE:EGFP(-) melanoma cells. This work identifies a TGFb induced immune response and demonstrates the need for the development of chronic TGFb biomarkers to predict patient response to TGFb inhibitors.}, keywords = {*Zebrafish *Melanoma/genetics/immunology/metabolism/pathology Animals Humans *Signal Transduction *Animals, Genetically Modified Green Fluorescent Proteins/metabolism/genetics Transforming Growth Factor beta1/metabolism Cell Line, Tumor Genes, Reporter Transforming Growth Factor beta/metabolism Gene Expression Regulation, Neoplastic TGFb cancer biology human macrophages melanoma zebrafish}, ISSN = {2050-084X (Electronic) 2050-084X (Linking)}, DOI = {10.7554/eLife.83527}, url = {https://www.ncbi.nlm.nih.gov/pubmed/38874379}, year = {2024}, type = {Journal Article} } @article{RN120, author = {Ostrem, B. E. L. and Dominguez-Iturza, N. and Stogsdill, J. A. and Faits, T. and Kim, K. and Levin, J. Z. and Arlotta, P.}, title = {Fetal brain response to maternal inflammation requires microglia}, journal = {Development}, volume = {151}, number = {10}, note = {Ostrem, Bridget Elaine LaMonica Dominguez-Iturza, Nuria Stogsdill, Jeffrey A Faits, Tyler Kim, Kwanho Levin, Joshua Z Arlotta, Paola eng R25 NS065743/NS/NINDS NIH HHS/ NS065743/NS/NINDS NIH HHS/ Stanley Center for Psychiatric Research, Broad Institute/ Harvard University/ NH/NIH HHS/ Klarman Cell Observatory, Broad Institute/ England 2024/05/22 Development. 2024 May 15;151(10):dev202252. doi: 10.1242/dev.202252. Epub 2024 May 22.}, abstract = {In utero infection and maternal inflammation can adversely impact fetal brain development. Maternal systemic illness, even in the absence of direct fetal brain infection, is associated with an increased risk of neuropsychiatric disorders in affected offspring. The cell types mediating the fetal brain response to maternal inflammation are largely unknown, hindering the development of novel treatment strategies. Here, we show that microglia, the resident phagocytes of the brain, highly express receptors for relevant pathogens and cytokines throughout embryonic development. Using a rodent maternal immune activation (MIA) model in which polyinosinic:polycytidylic acid is injected into pregnant mice, we demonstrate long-lasting transcriptional changes in fetal microglia that persist into postnatal life. We find that MIA induces widespread gene expression changes in neuronal and non-neuronal cells; importantly, these responses are abolished by selective genetic deletion of microglia, indicating that microglia are required for the transcriptional response of other cortical cell types to MIA. These findings demonstrate that microglia play a crucial durable role in the fetal response to maternal inflammation, and should be explored as potential therapeutic cell targets.}, keywords = {Animals *Microglia/metabolism/immunology Female Pregnancy Mice *Brain/pathology/immunology/metabolism *Inflammation/pathology/genetics *Poly I-C/pharmacology Fetus Mice, Inbred C57BL Gene Expression Regulation, Developmental Neurons/metabolism Cortical development Maternal immune activation Microglia Neuroinflammation}, ISSN = {1477-9129 (Electronic) 0950-1991 (Print) 0950-1991 (Linking)}, DOI = {10.1242/dev.202252}, url = {https://www.ncbi.nlm.nih.gov/pubmed/38775708}, year = {2024}, type = {Journal Article} } @article{RN119, author = {Pearce, S. and Lin, C. and Perez-Mercader, J.}, title = {Adaptive and Dissipative Hierarchical Population Crowding of Synthetic Protocells through Click-PISA under Gradient Energy Inputs}, journal = {Nano Lett}, volume = {24}, number = {8}, pages = {2457-2464}, note = {Pearce, Samuel Lin, Chenyu Perez-Mercader, Juan eng 2024/02/19 Nano Lett. 2024 Feb 28;24(8):2457-2464. doi: 10.1021/acs.nanolett.3c04035. Epub 2024 Feb 19.}, abstract = {The ability of living objects to respond rapidly en masse to various stimuli or stress is an important function in response to externally applied changes in the local environment. This occurs across many length scales, for instance, bacteria swarming in response to different stimuli or stress and macromolecular crowding within cells. Currently there are few mechanisms to induce similar autonomous behaviors within populations of synthetic protocells. Herein, we report a system in which populations of individual objects behave in a coordinated manner in response to changes in the energetic environment by the emergent self-organization of large object swarms. These swarms contain protocell populations of approximately 60 000 individuals. We demonstrate the dissipative nature of the hierarchical constructs, which persist under appropriate UV stimulation. Finally, we identify the ability of the object populations to change behaviors in an adaptive population-wide response to the local energetic environment.}, keywords = {Humans *Artificial Cells Macromolecular Substances artificial life click-PISA chemistry dissipative self-assembly ensemble behavior origins of life protocells}, ISSN = {1530-6992 (Electronic) 1530-6984 (Print) 1530-6984 (Linking)}, DOI = {10.1021/acs.nanolett.3c04035}, url = {https://www.ncbi.nlm.nih.gov/pubmed/38373157}, year = {2024}, type = {Journal Article} } @article{RN118, author = {Peter, M. and Shipman, S. and Heo, J. and Macklis, J. D.}, title = {Limitations of fluorescent timer protein maturation kinetics to isolate transcriptionally synchronized human neural progenitor cells}, journal = {iScience}, volume = {27}, number = {6}, pages = {109911}, note = {Peter, Manuel Shipman, Seth Heo, Jaewon Macklis, Jeffrey D eng DP1 NS106665/NS/NINDS NIH HHS/ R01 NS045523/NS/NINDS NIH HHS/ R01 NS049553/NS/NINDS NIH HHS/ R01 NS104055/NS/NINDS NIH HHS/ 2024/05/24 iScience. 2024 May 7;27(6):109911. doi: 10.1016/j.isci.2024.109911. eCollection 2024 Jun 21.}, abstract = {Differentiation of human pluripotent stem cells (hPSCs) into subtype-specific neurons holds substantial potential for disease modeling in vitro. For successful differentiation, a detailed understanding of the transcriptional networks regulating cell fate decisions is critical. The heterochronic nature of neurodevelopment, during which distinct cells in the brain and during in vitro differentiation acquire their fates in an unsynchronized manner, hinders pooled transcriptional comparisons. One approach is to "translate" chronologic time into linear developmental and maturational time. Simple binary promotor-driven fluorescent proteins (FPs) to pool similar cells are unable to achieve this goal, due to asynchronous promotor onset in individual cells. We tested five fluorescent timer (FT) molecules expressed from the endogenous paired box 6 (PAX6) promoter in 293T and human hPSCs. Each of these FT systems faithfully reported chronologic time in 293T cells, but none of the FT constructs followed the same fluorescence kinetics in human neural progenitor cells.}, keywords = {Biochemistry Biological sciences Cell biology Cellular neuroscience Molecular biology Natural sciences Neuroscience}, ISSN = {2589-0042 (Electronic) 2589-0042 (Linking)}, DOI = {10.1016/j.isci.2024.109911}, url = {https://www.ncbi.nlm.nih.gov/pubmed/38784012}, year = {2024}, type = {Journal Article} } @article{RN116, author = {Velayutham, N. and Garbern, J. C. and Elwell, H. L. T. and Zhuo, Z. and Ruland, L. and Elcure Alvarez, F. and Frontini, S. and Rodriguez Carreras, Y. and Eichholtz, M. and Ricci-Blair, E. and Shaw, J. Y. and Bouffard, A. H. and Sokol, M. and Mancheno Juncosa, E. and Rhoades, S. and van den Berg, D. and Kreymerman, A. and Aoyama, J. and Hofflin, J. and Ryan, H. and Ho Sui, S. and Lee, R. T.}, title = {P53 Activation Promotes Maturational Characteristics of Pluripotent Stem Cell-Derived Cardiomyocytes in 3-Dimensional Suspension Culture Via FOXO-FOXM1 Regulation}, journal = {J Am Heart Assoc}, volume = {13}, number = {13}, pages = {e033155}, note = {Velayutham, Nivedhitha Garbern, Jessica C Elwell, Hannah L T Zhuo, Zhu Ruland, Laura Elcure Alvarez, Farid Frontini, Sara Rodriguez Carreras, Yago Eichholtz, Marie Ricci-Blair, Elisabeth Shaw, Jeanna Y Bouffard, Aldric H Sokol, Morgan Mancheno Juncosa, Estela Rhoades, Seth van den Berg, Daphne Kreymerman, Alexander Aoyama, Junya Hofflin, Jens Ryan, Herb Ho Sui, Shannan Lee, Richard T eng England 2024/06/27 J Am Heart Assoc. 2024 Jul 2;13(13):e033155. doi: 10.1161/JAHA.123.033155. Epub 2024 Jun 27.}, abstract = {BACKGROUND: Current protocols generate highly pure human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) in vitro that recapitulate characteristics of mature in vivo cardiomyocytes. Yet, a risk of arrhythmias exists when hiPSC-CMs are injected into large animal models. Thus, understanding hiPSC-CM maturational mechanisms is crucial for clinical translation. Forkhead box (FOX) transcription factors regulate postnatal cardiomyocyte maturation through a balance between FOXO and FOXM1. We also previously demonstrated that p53 activation enhances hiPSC-CM maturation. Here, we investigate whether p53 activation modulates the FOXO/FOXM1 balance to promote hiPSC-CM maturation in 3-dimensional suspension culture. METHODS AND RESULTS: Three-dimensional cultures of hiPSC-CMs were treated with Nutlin-3a (p53 activator, 10 muM), LOM612 (FOXO relocator, 5 muM), AS1842856 (FOXO inhibitor, 1 muM), or RCM-1 (FOXM1 inhibitor, 1 muM), starting 2 days after onset of beating, with dimethyl sulfoxide (0.2% vehicle) as control. P53 activation promoted hiPSC-CM metabolic and electrophysiological maturation alongside FOXO upregulation and FOXM1 downregulation, in n=3 to 6 per group for all assays. FOXO inhibition significantly decreased expression of cardiac-specific markers such as TNNT2. In contrast, FOXO activation or FOXM1 inhibition promoted maturational characteristics such as increased contractility, oxygen consumption, and voltage peak maximum upstroke velocity, in n=3 to 6 per group for all assays. Further, by single-cell RNA sequencing of n=2 LOM612-treated cells compared with dimethyl sulfoxide, LOM612-mediated FOXO activation promoted expression of cardiac maturational pathways. CONCLUSIONS: We show that p53 activation promotes FOXO and suppresses FOXM1 during 3-dimensional hiPSC-CM maturation. These results expand our understanding of hiPSC-CM maturational mechanisms in a clinically-relevant 3-dimensional culture system.}, keywords = {*Myocytes, Cardiac/metabolism/drug effects *Forkhead Box Protein M1/metabolism/genetics *Tumor Suppressor Protein p53/metabolism/genetics Humans *Induced Pluripotent Stem Cells/metabolism/drug effects *Cell Differentiation Cell Culture Techniques, Three Dimensional/methods Cells, Cultured Signal Transduction Forkhead Box Protein O1/metabolism/genetics Foxm1 Foxo cardiomyocytes maturation p53 stem cells}, ISSN = {2047-9980 (Electronic) 2047-9980 (Linking)}, DOI = {10.1161/JAHA.123.033155}, url = {https://www.ncbi.nlm.nih.gov/pubmed/38934864}, year = {2024}, type = {Journal Article} } @article{RN124, author = {Walker, T. J. and Reyes-Alvarez, E. and Hyndman, B. D. and Sugiyama, M. G. and Oliveira, L. C. B. and Rekab, A. N. and Crupi, M. J. F. and Cabral-Dias, R. and Guo, Q. J. and Dahia, P. L. M. and Richardson, D. S. and Antonescu, C. N. and Mulligan, L. M.}, title = {Loss of tumor suppressor TMEM127 drives RET-mediated transformation through disrupted membrane dynamics}, journal = {Elife}, volume = {12}, note = {Wy1d9 Times Cited:0 Cited References Count:50}, abstract = {Internalization from the cell membrane and endosomal trafficking of receptor tyrosine kinases (RTKs) are important regulators of signaling in normal cells that can frequently be disrupted in cancer. The adrenal tumor pheochromocytoma (PCC) can be caused by activating mutations of the rearranged during transfection (RET) receptor tyrosine kinase, or inactivation of TMEM127, a transmembrane tumor suppressor implicated in trafficking of endosomal cargos. However, the role of aberrant receptor trafficking in PCC is not well understood. Here, we show that loss of TMEM127 causes wildtype RET protein accumulation on the cell surface, where increased receptor density facilitates constitutive ligand-independent activity and downstream signaling, driving cell proliferation. Loss of TMEM127 altered normal cell membrane organization and recruitment and stabilization of membrane protein complexes, impaired assembly, and maturation of clathrin-coated pits, and reduced internalization and degradation of cell surface RET. In addition to RTKs, TMEM127 depletion also promoted surface accumulation of several other transmembrane proteins, suggesting it may cause global defects in surface protein activity and function. Together, our data identify TMEM127 as an important determinant of membrane organization including membrane protein diffusability and protein complex assembly and provide a novel paradigm for oncogenesis in PCC where altered membrane dynamics promotes cell surface accumulation and constitutive activity of growth factor receptors to drive aberrant signaling and promote transformation.}, keywords = {ret tmem127 pheochromocytoma clathrin-mediated endocytosis membrane dynamics human clathrin protooncogene endocytosis receptor pheochromocytoma internalization susceptibility recruitment}, ISSN = {2050-084x}, DOI = {ARTN RP89100 10.7554/eLife.89100 10.7554/eLife.89100.3.sa1 10.7554/eLife.89100.3.sa2 10.7554/eLife.89100.3.sa3}, url = {://WOS:001258334200001}, year = {2024}, type = {Journal Article} }