The ecological state of the Persian or Arabian Gulf (hereafter ‘Gulf') is in sharp decline. Calls for comprehensive ecosystem-based management approaches and transboundary conservation have gone largely unanswered, despite mounting marine threats made worse by climate change. The region's long-standing political tensions add additional complexity, especially now as some Gulf countries will soon adopt ambitious goals to protect their marine environments as part of new global environmental commitments. The recent interest in global commitments comes at a time when diplomatic relations among all Gulf countries are improving. There is a window of opportunity for Gulf countries to meet global marine biodiversity conservation commitments, but only if scientists engage in peer-to-peer diplomacy to build trust, share knowledge and strategize marine conservation options across boundaries. The Gulf region needs more ocean diplomacy and coordination; just as critically, it needs actors at its science-policy interface to find better ways of adapting cooperative models to fit its unique marine environment, political context and culture. We propose a practical agenda for scientist-led diplomacy in the short term and lines of research from which to draw  to better design future science diplomacy practices and processes suited to the Gulf's setting. Classic case studies in science diplomacy—including the Antarctic Treaty 1959), the Outer Space Treaty 1967) and the creation of the Arctic Council (1996)—demonstrate how science engagement makes grander objectives possible . However, these examples of science diplomacy often reflect Western views of governance , conflic9, the Ou7 and the. 2As a restricted marine basin, the Gulf's maritime political boundaries are crowded among its neighbouring states of Bahrain, Iran, Iraq, Kuwait, Oman, Qatar, Saudi Arabia and the United Arab Emirates. During the summer, the Gulf is the world's hottest coastal environment. Many of its species live close to their physiological limits ,9. ClimaThe Gulf is also a global hotspot for rapid coastal development. Coastal reclamation, desalination plants and infrastructure projects are causing further declines in the Gulf's marine ecosystems  . BiodiveResponding to these common challenges across the region requires an integrative understanding of gaps in risk mitigation, hazard management and the magnitude of restoration measures needed to sustain marine species . RegionaPrevious calls for regional collaboration on ecosystem-based management (EBM) solutions have included coordinated ecosystem monitoring, transboundary fisheries governance, and a singular integrated conservation network of MPAs with other area-based conservation measures –19. Ther. 3The recent integration of regional cooperation on Gulf marine conservation began in the 1970s. Using the Mediterranean Action Plan as a template, the Regional Organisation for the Protection of the Marine Environment (ROPME) formed in 1978 among all Gulf coastal nations and marked early successes in controlling marine pollution. ROPME is affiliated with UNEP's Regional Seas Programme (RSP). Although widely seen as achievements of environmental diplomacy in the twentieth century, many of the 18 RSPs affiliated with UNEP today, including ROPME, are challenged by weak compliance and geopolitical issues .Intergovernmental organizations and global non-governmental organizations (NGOs) have also facilitated environmental diplomacy. For example, the World Wide Fund for Nature (WWF) regional office in UAE has co-led Gulf coral reef conservation planning across the coastlines of Abu Dhabi and Qatar . Gulf coThere is an urgent need to develop an inclusive framework to address climate-adjacent environmental challenges, such as marine biodiversity decline. Recent ROPME-led convenings on climate change with scientists and political leaders  demonstrData sharing requires the stewardship of data for public benefit and building strong infrastructure, like data institutions. In the Gulf, ROPME could meet this institutional need as a multilateral organization. Another model for data diplomacy involves fostering neutral boundary organizations. The Transnational Red Sea Center (TRSC) demonstrates this approach on the western side of the Arabian Peninsula. The TRSC facilitates knowledge sharing about coral reef habitats across the eight nations bordering the Red Sea. It seeks to overcome historical, conflict driven isolation through the diplomatic neutrality of the Swiss Foreign Ministry, which plays a boundary-spanning role by funding and advising the centre, including its transboundary expeditions, with the goal of cooperative coral science and dialogue . We see . 4Science diplomacy builds common interests among allies and historical adversaries. But even flagship settings for science diplomacy  show that trust and scientist-diplomat engagement can decline or fall to geopolitics ,5. ScienFirst, regional funding for scientists interested in networked Gulf studies should be dramatically expanded. Currently, such funding is insufficient to meet the moment and nationally delimited; pan-Gulf integration is often accidental. The GCC has strong funding capabilities and explicit coordinating infrastructure. Also, many national-level funding programs could provide enhanced regional cooperation support (e.g. Kuwait Foundation for the Advancement of Sciences (KFAS), Qatar Foundation's Qatar National Research Fund (QNRF), Iran National Science Foundation (INSF)). Additional financing can come from foundations, the private sector and international organizations.Second, peer networks among scientists should be strengthened to increase trust through data sharing. Conservation planning aimed at geopolitically transboundary species, such as commercial fish species, sea turtles and marine mammals, represents a unique area for sharing data. Grassroots networks among scientists are already building regional capacity for sharing data with global networks, as demonstrated by the ongoing engagement between the Arabian Seas Whale Network and the International Union for Conservation of Nature's Important Marine Mammal Areas process . ROPME cThird, scientists in the Gulf can lead transparent, participatory and systematic approaches to area-based conservation measures, which will accelerate progress towards global biodiversity conservation targets. Creating a comprehensive and ecologically coherent MPA network will provide an EBM strategy for enhancing the resilience of regional biodiversity in the face of climate change. We recommend that scientists, alongside capable boundary spanning organizations, help co-develop coordinated management metrics across MPAs to ensure effective conservation and connectivity . The GulOne of the problems with studies on marine ecosystems in the Gulf is the lack of integration in research topics. This disconnection is partially attributable to a lack of knowledge about ongoing research projects in the regional countries that suffer from journal paywalls and publication fees especially for conservation and biodiversity publications. To resolve this problem, we advocate creating a project database as a first step, which can be established and managed by the ROPME. A second solution is to create a forum such as Coral-List, which provides a forum for online discussion and announcements pertaining to coral reef ecosystem research, conservation and education. Having a forum for the researchers in the Gulf may help define integrative research topics that bridge the gap of information on marine ecosystems in the region. Identifying gaps will be especially useful for planning and coordination across all GCC member states and other pan-Gulf transboundary organizations. For example, the establishment of a network of Gulf MPAs requires understanding fundamental biological processes  among a large number of marine ecosystems across all member states.Scholars have long held-up less populated regions, like the Arctic, as settings for studying and advancing science diplomacy, while heavily populated and politically complex regions, like the Gulf, have been overlooked . We arguIn addition to increased diplomacy in the region, we call for better practices and processes for ocean diplomacy. This must be informed by relevant lines of social science scholarship—for example, the field of knowledge co-production, which captures the body of scholarship organized around ‘science-policy interface(s)' and other related concepts. Co-production has deepened scholars' and practitioners' understanding of the role of trust and boundary spanning organizations in informing policy-making –37. MultThe declining Gulf marine environments and the aligning of environmental interests, coupled with renewed regional diplomacy efforts, create an unusual convergence of drivers to begin understanding new modes of engagement. This window of opportunity for Gulf scientists to voluntarily integrate processes, data and strategize conservation research is potentially narrow, or at least time-sensitive relative to climate goals. Gulf governments, especially those states committing to ambitious 30 × 30 conservation targets , can tak


Information on how 3D genome topology emerged in animal evolution, how stable it is during development, its role in the evolution of phenotypic novelties and how exactly it affects gene expression is highly debated. So far, data to address these questions are lacking with the exception of a few key model species. Several gene regulatory mechanisms have been proposed, including scenarios where genome topology has little to no impact on gene expression, and vice versa. The ancient and diverse clade of spiralians may provide a crucial testing ground for such mechanisms. Sprialians have followed distinct evolutionary trajectories, with some clades experiencing genome expansions and/or large-scale genome rearrangements, and others undergoing genome contraction, substantially impacting their size and organisation. These changes have been associated with many phenotypic innovations in this clade. In this review, we describe how emerging genome topology data, along with functional tools, allow for testing these scenarios and discuss their predicted outcomes. Spiralia separated from the other major protostome lineage Ecdysozoa approximately 550 MYA , and is From an evolutionary perspective, one of the most fascinating aspects of spiralians is the diversity of their genomes. Ranging from the very small genomes of often parasitic or symbiotic species , including promoters, enhancers, silencers and operators, control the transcription of neighbouring genes. While some CREs are in close proximity to the genes they regulate, many are located further away and require precise folding of chromatin in order to perform their function. Chromatin interactions can be examined using assays for open chromatin such as ATAC-seq (assay for transposase-accessible chromatin) and multiomic (a combination of scATAC-seq and scRNA-seq) approaches  have rer detail , 57, ther detail , and absr detail ; therefor detail .HoxD locus in the mouse [Genome expansion may have distinct effects on genome topology and gene regulation. For example, when TADs expand correlatively with the genome size, novel interactions will only happen due to new regulatory sequences emerging or translocating into the TAD . The althe mouse , where the mouse .Dicyema japonicum [Dimorphilus gyrociliatus [HoxD-like ‘biphasic’ (or even ‘multiphasic’) regulatory interactions. Another possibility, given the very small genome size of these species, could also be the ‘disappearance’ of pervasive topology (due to CTCF loss), as is well known for the nematode Caenorhabditis elegans [The opposite force of genome expansion is DNA loss and subsequent genome contraction. Several spiralian genomes have undergone genome contraction compared with either the ancestral state, for example, the mesozoan aponicum  or the aciliatus  . In suc elegans . While nThe highly diverse and dynamic genome sizes, especially in spiralians, may play a key role in influencing 3D chromatin organisation which, in turn, will define the regulatory interactions and the constraints of their expansion or contraction over the macro-evolutionary time-scale. While alterations to TADs due to translocations have been extensively discussed , this imTo further test these observations, several state-of-the-art techniques can be applied. In particular, the recent application of multiomic approaches can find correlated open chromatin regions with target genes. This opens up avenues to test whether the correlated putative regulatory regions are in the direct vicinity, closer to the opposite TAD border or uncorrelated with the potential target gene. In addition, emerging tools on 3D modelling (e.g. ) are cruSpiralia includes many taxa that are highly diverse in their developmental or regenerative capabilities, such as planarian and annelid species that can regenerate every part of the body , as wellD. japonicum yielded just four Hox genes in this species which are also dispersed throughout the genome [Notospermus geniculatus is thought to lack a Hox cluster; however, this could be an artefact of a scaffold-level genome assembly [The diversity in both content and genomic linkage of the spiralian Hox genes is thought to contribute to their divergent body plans , 81. Note genome , 84. To assembly  and tranassembly , 87.Helobdella robusta reported in [D. gyrociliatus . Even tciliatus  has a coE. scolopes has very large intergenic distances (1 to 2.6 Mbp) in its Hox cluster, which are the largest known inter-Hox gene separators [The large differences in the spiralian genome sizes may put different emphasis on the role of genome topology in developmental gene regulation. For example, the cephalopod mollusc parators . Intergeparators . Regulatparators , 28. Recparators , 94 highTogether, new high-resolution chromatin interaction data will help resolve crucial developmental gene regulation in spiralians. Long-range interaction data, including inter-chromosomal contacts, may also shed light on whether spiralian chromosomes show evolutionarily conserved chromosomal-scale interactions as has been proposed recently using 3D modelling approaches . The obsE. scolopes uncovered hundreds of microsyntenies linked to coleoid cephalopod-specific traits such as their complex nervous system. HiC data revealed that these locally conserved gene neighbourhoods were often located in TADs [Drosophila suggests [Drosophila [Spiralian evolution has resulted in the evolution of a wide range of clade-specific adaptations, as encapsulated by their huge morphological and behavioural diversity. This makes the clade a perfect system in which to study the effect of genome topology on gene regulation and function. The reason/driver behind such fast pace remains to be characterised. Several studies suggest that reshuffling of TADs can ‘rewire’ CREs to act on alternate genes, resulting in transcriptional changes. Although compelling evidence of this exists in some species , evidenc in TADs . Further in TADs , though  in TADs . It shousuggests . Furtherosophila  and vertosophila .As well as through TAD rearrangements, genome topology may play a role in adaptation via more subtle changes in chromatin folding , e.g. thRecently, key papers described evolutionary breakpoints (EBR) in closely related vertebrate species , 107. AmSeveral closely related spiralian species can provide for excellent opportunities to evaluate the impact of EBRs on phenotypic diversity and for understanding the effect of genome size on EBRs. Recently, several such genomes within bivalves , 110, anTo conclude, it remains to be seen how significant genome topology data will be in unravelling the different modes of gene regulation in spiralians. In this review, we describe how this role will likely depend on the genome size and gene regulatory modality resulting from the species’ evolutionary history. The plethora of phenotypic traits that emerge in spiralians and the diversity of the underlying developmental programmes has provided for fruitful ground for many evo-devo studies. The addition of more high-quality reference genomes and 3C data to studies such as these can reveal whether genome topology puts increased emphasis on distal enhancers in gene regulation or how neighbouring TADs can contribute to gene regulation at TAD boundaries. As genome-wide methods for genome topology assessment as well as functional tools such as CRISPR-Cas9, and high-resolution expression studies such as multiomics, become readily available, spiralians will likely develop into valuable emerging systems for the study of the role of genome topology in their evolution.In addition, as more chromosomal-scale genomes including genome topology data become available for this clade, the investigation into the variety of molecular mechanisms can take off. Transposable element-driven genome size differences in closely related species of molluscs, polychaetes, as well as whole genome duplications and rearrangements e.g. in cephalopods have had profound effects on their genomes and topology. So far, clear genome topological structures exist in the few spiralian  genomes where this has been investigated. While the general genome topology and compartmentalisation (i.e. TAD presence) looks similar to other bilaterian genomes, how those structures are maintained during genome expansion, contraction, across development and whether they change between tissues or cell types will be a crucial emerging topic in the next few years. Taken together, insights into the function of genome topology from spiralians may be crucially informative not only in the understanding of their evolution, but also in the function of topology in well-established model organisms.Spiralians comprise one of the most species-rich, morphologically and genomically diverse clade of animals.While ample progress has been made at establishing several spiralian model systems, little is known about how their genomic organisation may impact gene regulation.This review discusses recent progress in the field of spiralian genome evolution and topology, and outlines several testable predictions for the effect of genome topology on gene regulation in spiralians.In particular, the review proposes that the often very dynamic genome expansion and contraction has played a central role in the emergence, modification and degradation of topologically associating domains (TADs) in spiralian genomes and species-specific adaptations.


To fulfil the growing needs of the global population, sustainability in food production must be ensured. Insect pests and pathogens are primarily responsible for one-third of food losses and harmful synthetic pesticides have been applied to protect crops from these pests and other pathogens such as viruses and fungi. An alternative pathogen control mechanism that is more “friendly” to the environment can be developed by externally applying double-stranded RNAs (dsRNAs) to suppress gene expression. However, the use of dsRNA sprays in open fields is complicated with respect to variable efficiencies in the dsRNA delivery, and the stability of the dsRNA on and in the plants, and because the mechanisms of gene silencing may differ between plants and between different pathogen targets. Thus, nanocarrier delivery systems have been especially used with the goal of improving the efficacy of dsRNAs. Here, we highlight recent developments in nanoparticle-mediated nanocarriers to deliver dsRNA, including layered double hydroxide, carbon dots, carbon nanotubes, gold nanoparticles, chitosan nanoparticles, silica nanoparticles, liposomes, and cell-penetrating peptides, by review of the literature and patent landscape. The effects of nanoparticle size and surface modification on the dsRNA uptake efficiency in plants are also discussed. Finally, we emphasize the overall limitation of dsRNA sprays, the risks associated, and the potential safety concerns for spraying dsRNAs on crops. As the world population is expected to reach 9.8 billion by 2050 , global Scientists have explored RNAi, which is a specific post-transcriptional cellular mechanism, as an alternative tool for crop protection strategies. RNAi technology allows the manipulation of regulatory mechanisms to silence genes in plant and animal cells using sequence-specific, small double-stranded RNAs (dsRNAs). Mimicking a laser-guided mechanism in precision, dsRNAs are produced by copying complementary gene sequences of pests and pathogens, and these dsRNAs can be used as a template to destroy pests’ and pathogens’ mRNA sequences, ultimately disrupting protein production . TransfoAs an alternative to the transgenic approach, scientists have explored the use of naked dsRNAs, which can be applied directly onto plants without inserting plasmid DNAs. Also referred to as spray-induced gene silencing (SIGS), the first report of exogenous dsRNAs in plants was carried out by mechanically inoculating bacterially produced naked dsRNAs on pepper mild mottle virus (PMMoV) infected plants to induce RNAi effects for the triggering step of post-transcriptional gene silencing, thereby targeting viral RNAs for degradation . BesidesAnother important challenge for spray-on applications of dsRNAs is the unique barriers of walled plant cells. The plant cell wall is composed of a tough semipermeable matrix that serves not only as a protective barrier to the external environment but also as a barrier to the uptake of foreign materials into plant cells . While tOne possible approach to solving challenges associated with impaired endosomal escape, dsRNA degradation and poor cellular uptake of dsRNA is by using nanocarriers as a delivery vehicle for dsRNAs . NanocarRecent publications on non-transformative RNAi strategies were reviewed by searching the Thompson Reuters Web of Science (WoS) database using combinations of search terms (Queries), asterisks and Boolean operators. Queries were as follows: (1) TS =  AND TS = (*RNA silencing); (2) TS =  AND TS = (*RNA*) AND TS = (crop); (3) TS =  AND TS = (*RNA*) AND TS = (plant). This selection was further refined to include research articles only that were published between 2017 and 2022. The initial search found 1631 journal publications. Next, duplicated entries were removed, and abstracts of the papers were reviewed manually to filter publications that met two inclusion criteria: (1) research articles and (2) methodological approach that uses nanoparticles to deliver RNAs into plant cells. The filtering process resulted in 16 relevant publications that experimentally investigated potential applications of topical dsRNA sprays on crops.These publications described seven novel designs of nanocarriers (delivery vehicles) for RNAs into plant cells. Important details, such as nanoparticle design, RNA silencing activity and mode of delivery are presented in 2+1−xM3+ x(OH)2][An−]x/n·zH2O, where M2+ and M3+ are divalent and trivalent metal ions, and An– is the interlayer charge-balancing anion. A great amount of work has demonstrated the multi-purpose nature of LDH including as a biocompatible, low-toxic transporter for gene and drug delivery in mammalian cells [Layered Double Hydroxides LDH,  are hydran cells . More rean cells ,25,26,36The uptake of LDH by plant cells has been reported as both by free penetration as well as by clathrin-mediated-endocytosis (CME), in which the LDH nanosheets are engulfed by clathrin-coated vesicles before they are released and dispersed within cytosols . As the Methods of delivery for long dsRNAs encapsulated by LDH nanosheets were shown to influence RNAi efficiency against a target. For example, spraying of dsRNAs-LDH resulted in the highest reduction of fungal disease severity compared to leaf petiole adsorption and root dipping . High-prAlthough LDH uptake was reported in several plant parts including leaves ,26 and pMuch interest in carbon dots CDs, a in planCDs are also versatile as a delivery vehicle of genetic materials into plant cells. A recent study reported that amine-functionalized CD (CD-PEI) is suitable as a nanocarrier for siRNA to silence transgenes and endogenous plant genes . Using aCarbon nanotubes CNTs,  are cyliSubsequent studies have elucidated the ability of CNT as a carrier of other biomolecular cargoes into intact plant cells, for example, cellulase , plasmid2 is protonated and forms -NH3+. The positive charges elicit electrostatic interactions with negatively charged phosphate backbones of nucleic acids when mixed in solution, and genetic materials are encapsulated in stable nanostructures (nanoparticles) for efficient gene delivery [Chitosan  is a deadelivery .While there is a larger body of research on the use of chitosan carriers in animal cells, which have shown good protection against nucleases ,53,54, aTribolium castaneum and Acyrthosiphon pisum, consequently impairing protein folding and resulting in premature death of these plant pests. More importantly, ingestion of BPAC-dsRNA showed a lethality rate of 6 to 9 days earlier compared to feeding with dsRNAs alone, suggesting BPAC complexation enhanced oral delivery of dsRNAs and resulted in improved RNAi effects.The topical application of peptides as the nucleic acid carrier has been one of the major focuses for delivery in plants. Peptides are short chains of amino acids that carry positive charges on the cationic groups. The positive charges interact electrostatically with negatively charged nucleic acids to encapsulate the genetic materials for an efficient delivery ,55,56. P9 allows the CPPs to have a greater functional presence at the surface of the genetic material-CPP complexes [In plant cells, functionalizing cell-penetrating peptides CPP, , for exaomplexes . The desNicotinum benthamiana while being non-toxic to the host plant.Gold (Au) refers to the solid inorganic aurum metal nanoparticle that has been used in various molecular delivery applications ,43. Au nSilica nanoparticles SNP,  have proLiposomes  are surfAn important aspect of topical RNAi application is the need for risk assessments and management of the dsRNA-based products. Several key aspects for consideration for risk assessments have been described in reports such as the US EPA’s “White paper on RNAi technology as a pesticide: Problem formulation for human health and ecological risk assessment”  and the http://www.flyrnai.org; accessed on 29 November 2022) [http://www.genomernai.org; accessed on 29 November 2022) [http://dscheck.rnai.jp; accessed on 29 November 2022) [https://www.specifly.org; accessed on 21 December 2022) [https://github.com/snowformatics/siFi21; accessed on 29 December 2022) [https://www.zhaolab.org/pssRNAit; accessed on 26 December 2022) [https://github.com/pfred; accessed on 21 December 2022) [Bioinformatics tools and models have been particularly useful in designing RNAi targets and predicting potential off-target predictions. Reliable searching, predictions and designs of RNAi triggering sequences were made possible with the availability of genomic data libraries of numerous species such as the Drosophila RNAi Screening Center (available at er 2022)  and Genoer 2022) , web-baser 2022)  and Offter 2022) , as weller 2022) , pssRNAier 2022)  and PFREer 2022) . Nonetheer 2022) . The seler 2022) .C. elegans was reported when chitosan was used as either polyplex nanoparticles or alone [There also remain uncertainties over the possible fates of dsRNAs once they are translocated into the target cells. For example, considering clathrin-dependent endocytosis is a highly conserved mechanism across eukaryotic species , there ior alone . Thus, aLeptinotarsa decemlineata (submitted by Syngenta) [The commercial potential for RNAi technologies could be hampered by uncertainties in biosafety regulatory pathways for the technology. While many other countries are yet to clarify their regulatory positions for RNAi sprays, several countries have made the first move in reviewing and defining their regulatory processes. In the USA, the risks of RNAi-based biopesticides are evaluated using chemical pesticide templates while Canada, which adopts a product-based biosafety regulation, oversees topically applied RNAi  based on trait novelty. Australia also has taken a favorable standpoint towards regulating non-transgenic RNAi and approved the proposal to exempt topically applied dsRNAs from GMO regulations (refer to schedule 1A techniques that are not gene technology), while New Zealand ruled that dsRNA-treated eukaryotic organisms do not meet the definition of a GMO (APP203395) ,87. Suchyngenta) . Both fiIt is also important to note that one of the commercialization barriers to topical RNAi applications are the high expense of large-scale dsRNA production, especially if the synthesis is carried out using in vitro transcription kits. Private startup firms, for example, GreenLight Biosciences  and RNAgri , have developed and patented proprietary dsRNA-mass production methods to lower the cost . AlternaThe topical application of dsRNA as active molecules presents a highly versatile crop management strategy that does not require plant transformation methods. Since naked dsRNAs are susceptible to degradation upon exposure to the environment, dsRNAs can be encapsulated or layered with biocompatible nanoparticles to prolong the RNA stability and increase silencing efficiency when applied in field-like conditions. However, the design and selection of RNA carriers will depend on several factors such as the RNA size , the target  and the method of delivery .This work reviewed a selection of nanocarrier delivery systems and their use, such as LDH, CD, CNT, gold nanoparticles, chitosan nanoparticles, silica nanoparticles, liposomes and CPP. The use of these nanocarriers allows dsRNAs to be more stable and efficient when applied in open fields. In perspective, the nanocarriers must first be surface-modified with polar components to allow for carrier-RNA polar interactions. Further studies are also needed to understand the possible toxicity effects of dsRNA-nanocarrier complexes and how various barriers in the cellular system influence the uptake, silencing effects and systemic protection mechanisms in plants.Despite the promising potential of topical RNAi technology, there are limitations, potential risks and safety concerns in relation to the technology that need to be addressed. Potential off-target effects, uncertainties in the fate of dsRNAs and regulation of the technology, as well as the high production cost of the dsRNAs, may limit the commercial potential of the technology. More importantly, good governance of topical RNAi technology demands a greater corporate responsibility that requires continuous dialogues with relevant stakeholders and that devotes more attention towards addressing ethical issues and societal costs of the technology. To ensure the longevity of topical RNAi application, the authors believe that understanding society’s position on the technology with respect to the aspects of willingness-to-pay, technological adoption trade-offs and public trust are key areas that need urgent attention.


Vaccines have significant potential in treating and/or preventing diseases, yet there remain challenges in developing effective vaccines against some diseases, such as AIDS and certain tumors. Mesenchymal stem cells (MSCs), a subset of cells with low immunogenicity, high proliferation potential, and an abundant source of extracellular vesicles (EVs), represent one of the novel and promising vaccine platforms. This review describes the unique features and potential mechanisms of MSCs as a novel vaccine platform. We also cover aspects such as the safety and stability of MSCs that warrant future in-depth studies. Vaccines refer to biological products made by various pathogenic microorganisms for vaccination. Traditionally, vaccines have played essential roles in preventing infectious diseases ,2. Ever Viral vectors have been widely used. There are replicating viral vectors and non-replicating viral vectors, also known as inactivated vaccines and live attenuated vaccines . InactivThe proliferation rate of the live attenuated vaccine is inhibited, and the slower replication rate avoids the body’s overreaction to the vaccine, but is sufficient to stimulate the immune system to respond and form immune memory. However, the purification of live attenuated vaccines takes a long time, through the continuous selection of virus strains with reduced virulence, and cannot be too weak to activate the immune system ,12. WithSubunit vaccines take the representative antigens of the whole pathogen as the main body, which can be selected from the intact pathogen, or can be produced through genetic recombination engineering . RepreseThe protein vaccine is currently the most widely used type of vaccine. Diseases including influenza, hepatitis B, and malaria can now be effectively prevented by protein vaccines . But theThe exotoxins of some pathogens can cause disease. Toxoid vaccines use inactivated toxins as antigens to stimulate immune responses. The toxoid is not pathogenic, but retains its ability to activate the immune system . The bigCapsular polysaccharides of some pathogens can also be used as antigens for subunit vaccines. However, the immunogenicity of the polysaccharide vaccine is very poor, only providing short-term protection, and the capsular polysaccharide can be directly recognized by B cells without the help of T cells, so it is largely ineffective for children with immature immune systems .Although the above several types of vaccines have made great progress for human health, they still have various deficiencies and side effects, such as low efficacy and reversible toxicity. New vaccine types still need more and more in-depth research . Below wNucleic acid vaccines represent another novel vaccine platform. They are currently the most likely field for humans to invent universal vaccines. The DNA vaccine expresses the immunogen of interest upon being translocated into the nucleus of specific host cells . As a reAlthough the above several types of vaccines have made great progress for human health, they still have various deficiencies and side effects, such as live attenuated vaccines’ possible reversible toxicity and DNA vaccines’ weak immunogenicity . One vacWith the gradual research on the various characteristics of MSCs, there is hope for new therapies for a variety of incurable diseases. Much research is optimistic about the potential of MSCs as the new generation of vaccines platform. MSCs are widely distributed in almost all tissues (including fetuses and adults), such as bone marrow, blood, umbilical cord blood, placenta, fat, amniotic fluid, dental pulp, skin, menstrual blood, etc. ,26. EndoAs a poorly differentiated naive cell, the MSC has the potential for multi-directional differentiation and self-replication, and is in an undifferentiated dormant state in the local microenvironment of the body . Once thThe low immunogenicity of MSCs comes from the fact that MSCs do not express major histocompatibility complex II(MHC II) molecules and only express low-level MHC I molecules. Therefore, MSCs have the characteristics of immune exemption, and they can barely stimulate immune reaction and avoid the attack of toxic T cells and NK cells ,35. AfteEVs are microvesicles secreted by cells. They can be divided into various subtypes according to different secretion mechanisms, but it is difficult to separate specific subtypes. They are usually divided according to their particle size into small (30–150 nm), medium (100–1000 nm), and large EVs (50–5000 nm). EVs are closely related to tissue homeostasis, and carriers of intercellular communication. They perform a bidirectional role in regulating human physiological and pathological processes. EVs are involved in almost all physiological activities, including cell proliferation, migration, and apoptosis. EVs are deeply involved in immune surveillance and activation of immune cells ,42. MSC-The MSC shows excellent superiority as a vaccine material for treating and preventing diseases. Meanwhile, the possibility of the MSC as a vaccine carrier deserves more attention as a new type of vaccine with self-replication, multi-directional differentiation potential, and easy to obtain and abundant EVs sources. It has been reported that EV products derived from bone marrow mesenchymal stem cells were safe and effective in the treatment of severe COVID-19. The detailed mechanism of MSCs inducing or suppressing immune cells needs further study. All immune cells involved in the inflammatory process can secrete EVs, and they play multiple roles in the inflammatory process. In the immune response, EVs are involved in activating the immune system and stimulating immune cell differentiation, mainly through the following three mechanisms: (1) Simple fusion of cell membranes, (2) Antigen endocytosis of immune cells, (3) Activation by specific membrane surface antigen recognition. The anti-inflammatory tendency of MSC-EVs increases the targeting of EVs to infected endothelial cells and immune cells. After contact with the recipient cell, the EVs membrane protein antigens TIM1 and TIM4 link to the membrane receptor of the recipient cell and finally complete the membrane fusion. The contents are then released into the cytoplasm, resulting in the change in the recipient intracellular environment. EVs combine the advantages of synthetic nano carriers and cell-mediated carriers, and are easily membrane-modified to improve their targeting ability and specificity ,48,49. TCancer-cell-derived EVs can rapidly activate the immune system response because they carry tumor-specific antigens on their surface that are quickly recognized by APC and thus inhibit in situ and metastatic cancer cells. The uptake efficiency of EVs by tumor cells is better than that of synthetic nanoparticles. Circular RNA is a potential gene delivery therapy, but is often trapped in endosomes. Strategies that combine EVs with SNAs can bypass these limitations, and SNA-EVs are significantly more efficient at silencing miRNAs than naked SNAs. EVs-mediated mRNA expression drives functional enzyme expression in vivo, and is more efficient than lipid nanoparticle formulations. The use of liposomes has been reported to lead to adverse immunogenic responses, which can be effectively avoided by extracellular vesicle carriers ,51.Traditionally, vaccines work by harnessing the immune system’s ability to recognize specific markers of pathogens, creating lasting protective effect. For the COVID-19 pandemic that is raging around the world, no effective preventive measures have been found temporarily. In this situation, the development of targeted vaccines is imminent. Although humans have developed effective vaccines for a variety of serious infectious diseases, the variability of viral nucleic acids makes vaccine development still difficult, and many diseases still lack effective vaccine protection, such as HIV and malaria ,53. TherThe MSC is a unique pluripotent stem cell and is currently used as a gene therapy vector for the treatment of various diseases including cancer and autoimmune diseases . The natThe vaccine function of MSCs cultured in vitro is mainly realized by the expression of target antigens by transfected exogenous genes, which can express the antigen protein of human tumors and the surface antigens of external pathogens ,59. And The accurate recognition of tumor-specific antigens and the escape of tumor cells are the biggest difficulties for the immune system in locating and quickly killing tumor cells . Tumor-aThe function of MSCs depends on cytokines secreted by the tumor immune cells in which they are located. Depending on differences in microenvironmental cytokines, MSCs can be transformed to promote and inhibit tumor phenotypes, thus altering the rate of tumor development . At the With the orienting properties of MSCs, MSCs and their EVs can deliver anti-tumor drugs to tumor cells, kill tumor cells, and inhibit the invasion of tumor cells. This is because MSCs have different chemotactic-related receptors, orienting to the tumor site and participating in the immune response against tumor cells. Studies have used GPC3+/CD3+ MSCS to induce CD3+ specific T cells to recognize GPC3+ cancer cells. The co-culture of the above three types of cells led to an increase in the production of IFN-γ by specific T cells in the microenvironment, and activated the killing effect of GPC3+ specific cytotoxic T lymphocytes(CTL), which led to the apoptosis of GPC3+ tumor cells. This phenomenon has also been demonstrated in animal studies, which suggest that MSCs are highly suitable for inducing tumor immunotherapy. MSCs have advantages as therapeutic tumor vaccine carriers ,67.Compared with other drugs, MSCs loaded with tumor drugs do not affect surrounding healthy tissue, but precisely target the primary and metastatic sites of tumors to release drugs. Therefore, drug-loaded MSCs treatment has fewer side effects than chemotherapy. In animal experiments, the half-life of modified MSCs was prolonged and the tumor inhibition effect was better. MSCs vaccine therapy can also work synergistically with chemotherapy. By carrying exogenous IL-2, engineered MSCs successfully overcome chemotherapy resistance caused by immune checkpoint blockade (ICB), activate CD8+ CTL in vivo, and induce systemic anti-tumor effects. The drugs carried by MSCs include not only small molecule drugs, miRNAs, etc., but also viruses that can kill tumors . MSCs caMSC-derived EVs have good therapeutic effects against different diseases and are strong candidates for MSC-based cell-free therapies. Several studies have shown that MSC-EVs contain tumor-inhibiting miRNAs expressed by MSCs and can be used as an MSC vaccine therapy for tumor immunotherapy. EVs have a bilayer lipid membrane that easily passes through the biological barrier in vivo and delivers miRNAs with anti-tumor effects directly to target cells. MSC-EVs can cross the blood–brain barrier (BBB), migrate to brain tumor cell sites, and exert anti-tumor effects by delivering drugs. Thus, MSC-EVs can deliver specific drugs directly into tumor cells, thereby inducing tumor cell apoptosis and inhibiting invasion .MSCs are regarded as a powerful new vaccine to prevent tumor occurrence and control tumor metastasis . The resTransfected human mesenchymal stem cells, which are like a small protein antigen response device, are a promising approach of anti-virus vaccines. The most important function of anti-virus vaccines is to induce adaptive immunity of sufficient strength, so that the human immune system can form a reliable immune response to susceptible pathogens . AntigenMSCs have the potential to become a new type of vaccine due to their unique immunomodulatory effect, differentiation potential, and abundant source of EVs. The MSC after gene transfection can express the surface antigen of the pathogen, simulate natural infection, and induce a specific immune response. EVs are another hopeful choice to express pathogenic antigen; cell-free therapy avoids the defects of heterologous gene integration into the genome of the host cell. After sufficiently reliable tests, the MSC vaccine will have considerable potential to treat currently incurable diseases. But the study of MSCs as a vaccine is still in the laboratory stage and has not yet undergone large-scale clinical trials. Therefore, its safety and efficacy still need to be further verified. It will take quite some time for MSCs to be marketed as a vaccine, but this does not affect our continuing in-depth research in this field.


Bupropion is an antidepressant utilized widely for the treatment of various mood disorders and smoking cessation due to its favorable side effect profile, cost, and response to therapy. While serious adverse reactions are rare, in the decades since its approval by the FDA, multiple cases of serum sickness-like reactions to bupropion have been reported, amongst other adverse drug reactions (ADRs). This report is of the case of a 25-year-old female who developed a serum sickness-like reaction to bupropion 21 days after initiation of treatment. She did not respond to conservative therapy but did respond promptly to oral corticosteroids and discontinuation of bupropion. This case serves to bolster the existing literature surrounding ADRs to bupropion and other antidepressant medications in the categories of systemic and dermatologic manifestations.  This case report describes the clinical presentation of a serum sickness-like reaction associated with a frequently prescribed atypical antidepressant medication (bupropion hydrochloride (HCl)), its diagnosis, and subsequent management. Bupropion is a norepinephrine/dopamine reuptake inhibitor that is commonly used to treat various mood disorders, most notably major depressive disorder and seasonal affective disorder. It can also be prescribed for smoking cessation, attention-deficit/hyperactivity disorder (ADHD), and obesity . While tOn September 9, 2021, a 25-year-old female with a medical history of generalized anxiety disorder and major depressive disorder presented to her primary care physician (PCP) for evaluation of the two-day history of rash, angioedema, and polyarthralgias. The patient reported she woke up in the early morning of September 8, 2021, with symmetrically grouped papules on her upper extremity distal extensor surfaces without any associated pruritis or pain Figures , 1B. HouDue to her worsening condition and failure of conservative treatment, the patient presented to her PCP for evaluation on September 9, 2021. After conducting a thorough history and physical, it was found that the patient was started on bupropion HCl 150 mg once daily 21 days before the onset of symptoms. She had never experienced similar symptoms, and the only recent lifestyle change she implemented was the addition of this antidepressant medication. As such, the patient was diagnosed clinically as experiencing an adverse reaction to the bupropion HCl. No laboratory studies or imaging tests were conducted due to the clinically diagnostic nature of drug-induced serum sickness-like reaction. For further evidence supportive of a drug-induced reaction, the Naranjo algorithm  was employed, resulting in a score of six. This score indicated the patient’s clinical presentation was probably the result of a drug reaction , topical corticosteroids, systemic corticosteroids if needed, and prompt discontinuation of bupropion. Prior case series have suggested that such reactions typically resolve within around 10 days of treatment .Cases of bupropion-induced serum sickness-like reactions have been reported previously. Serious adverse reactions (SARs) to bupropion were characterized in a French analysis of reported reactions since marketing authorization for bupropion in 2001. This analysis of reported adverse reactions over a three-year timeframe included 40 cases of serum sickness-like reactions out of an exposed population of 692,798 (< 0.006%) .Previously reported cases of bupropion-induced serum sickness-like reactions have demonstrated strikingly similar clinical presentations as described in this case. Notably, existing case reports have described marked improvement in systemic symptoms  within hours of initiation of treatment  -14. WhilWhile bupropion is generally a well-tolerated medication for various mood disorders, it has been known to cause delayed hypersensitivity serum sickness-like reactions in a small percentage of patients. Symptoms of this reaction can include pruritis, urticaria, angioedema, and polyarthralgias, which can be very concerning for patients given the sudden onset of symptoms and the potential for rapid evolution and failure of symptoms to resolve spontaneously. Discontinuation of the offending agent with the use of conservative measures such as oral antihistamines and topical steroid creams have been shown to aid in the resolution of such symptoms. However, refractory or severe cases can be managed with systemic corticosteroids. Clinicians may consider counseling patients regarding the possibility of this adverse reaction when initiating bupropion with prompt discontinuation and consideration of initiation of various other therapies for their underlying conditions if such a reaction occurs.


Neurodegenerative and neurovascular disorders affect millions of people worldwide and account for a large and increasing health burden on the general population. Thus, there is a critical need to identify potential disease-modifying treatments that can prevent or slow the disease progression. Mitochondria are highly dynamic organelles and play an important role in energy metabolism and redox homeostasis, and mitochondrial dysfunction threatens cell homeostasis, perturbs energy production, and ultimately leads to cell death and diseases. Impaired mitochondrial function has been linked to the pathogenesis of several human neurological disorders. Given the significant contribution of mitochondrial dysfunction in neurological disorders, there has been considerable interest in developing therapies that can attenuate mitochondrial abnormalities and proffer neuroprotective effects. Unfortunately, therapies that target specific components of mitochondria or oxidative stress pathways have exhibited limited translatability. To this end, mitochondrial transplantation therapy (MTT) presents a new paradigm of therapeutic intervention, which involves the supplementation of healthy mitochondria to replace the damaged mitochondria for the treatment of neurological disorders. Prior studies demonstrated that the supplementation of healthy donor mitochondria to damaged neurons promotes neuronal viability, activity, and neurite growth and has been shown to provide benefits for neural and extra-neural diseases. In this review, we discuss the significance of mitochondria and summarize an overview of the recent advances and development of MTT in neurodegenerative and neurovascular disorders, particularly Parkinson’s disease, Alzheimer’s disease, and stroke. The significance of MTT is emerging as they meet a critical need to develop a disease-modifying intervention for neurodegenerative and neurovascular disorders. Unfortunately, despite decades of research, there is no effective treatment available for these diseases. Thus, patients with neurological disorders are in urgent need of disease- modifying therapies that can slow or stop the progression of the neurodegenerative process. However, a major hurdle to the development of new therapies is the gap in our knowledge of the etiology of neurodegenerative and neurovascular disorders. The importance of the mitochondrion in the maintenance and preservation of neuronal survival and function is well established, and multiple lines of evidence suggest that mitochondrial dysfunction plays a key role in the pathogenesis of many neurodegenerative and neurovascular disorders such as Alzheimer’s disease (AD), Parkinson’s disease (PD), and ischemic stroke -4.1) [Mitochondria are membrane-bound cellular organelles which play an important role in the production of adenosine triphosphate (ATP) through oxidative phosphorylation (OXPHOS). However, OXPHOS represents a significant source of toxic endogenous free radicals as byproducts of normal cellular respiration. In addition to energy generation, mitochondria play a crucial role in a myriad of physiological functions, including fatty acids biosynthesis, calcium buffering, and integration of signaling pathways that educate immune response, autophagy, and cell survival -7. Howev1) -29. More1) , 30. MTT1) . The ear1) , and MTT1) -35. Find1) , 36, 37.Interestingly, MTT is a rapidly growing area of focus in the neuroscience research field. Its potential as a novel therapy is evident from the exponential increase in the number of published articles on MTT as searches that were completed on PubMed, MTT and neurological disorders showed 240 articles that were published in a period from 2000 to 2015. Evidently, MTT in neurological disorders has received more attention, with 267 articles on PubMed in the period from 2015 to 2022. These studies led to the emerging concept that MTT is a potential and feasible therapeutic approach to neurodegenerative and neurovascular disorders. However, MTT must still be explored for its safety and mechanism of therapeutic benefits. In this review, we assess mitochondrial dysfunction in neurodegenerative and neurovascular disorders and discuss the recent advances in MTT as a novel and promising treatment for neurodegenerative and neurovascular disorders.2Mitochondria are double-membrane cellular organelles and are often known as the powerhouse of the cell because they are the primary unit for generating ATP through OXPHOS. Although they are known to create energy for a cell to function, mitochondria also regulate multiple cellular processes involved in survival and death, lipid biogenesis, calcium buffering, cellular signaling, and degradation of misfolding proteins -7, 38. TMitochondrial dynamics play an important role in ensuring mitochondrial homeostasis and quality and are tightly regulated by the fusion and fission machinery. Mitochondria fusion is mediated by the GTPases Opa1 and Mitofusin-1 (Mfn1) and Mfn2, whereas fission is mediated by the mitochondrial fission 1 protein (Fis1), mitochondrial fission factor (Mff), and dynamin-related protein (Drp1). Impaired mitochondrial dynamics can lead to abnormal transport and distribution of mitochondria in neurons and can negatively impact synaptic and neuronal function . Mitopha2.1via modulating neurotransmitter disturbance. Decades of research on rodents and humans showed that mitochondrial dysfunction plays a significant role in AD [Alzheimer’s disease (AD) is the most common form of age-associated dementia. Nationwide, the incidence of AD and care costs is rising nearly exponentially. The medical cost for AD is expected to be $ 290 billion in 2019, and caregivers will provide an estimated 18.5 billion hours in unpaid care valued at $234 billion . AD is cle in AD -54. The le in AD . In supple in AD , 56, 57.Glucose utilization in the brain is broadly used as a primary measure to determine energy metabolism. PET imaging studies have demonstrated a significant decline in glucose utilization in brain regions involved in learning and memory of AD patients as compared to controls , 59. TheMitochondria develop a sophisticated mitochondrial quality control system to manage inevitable damage to its contents or superfluous mitochondria. At the organelle level, damaged mitochondria are degraded by a selective form of autophagy which is termed mitophagy. Growing evidence suggests that mitophagy function is impaired in AD, resulting in the accumulation of dysfunctional mitochondria and promotes Alzheimer’s related neuropathology and cognitive deficits , 81. Int2.2Parkinson’s disease (PD), a progressive neurodegenerative disease, is characterized by the death or malfunction of dopaminergic neurons in the substantia nigra and dopamine depletion in the striatum resulting in loss of motor functions , 84. Pat2.3Stroke is the most common cause of serious disability for adults and the leading cause of death worldwide. According to the World Health Organization (WHO), approximately 15 million people worldwide suffer from stroke each year. Of these, more than 5 million die and another 5 million are left permanently disabled, placing a huge burden on the family and the community . To dateDuring reperfusion, blood flow may be restored in time to maintain neuronal viability; however, reperfusion can itself induce mitochondrial to produce substantial amounts of ROS . MitochoThe B-cell lymphoma (BCL-2) protein family, a major regulator of the outer mitochondrial membrane, plays an important role in the regulation of neuronal death in ischemic stroke , 123. Ad3via pharmacological compounds or genetic approach are potential therapeutic approaches which render neuroprotective effects. For example, Drp1 inhibition reduces mitochondrial fission and preserves normal mitochondrial morphology and function during neurological conditions [Mitochondrial medicine refers to disease-modifying intervention for diseases or conditions caused by mitochondria dysfunctional. Mitochondrial medicine attempts to mitigate cellular dysfunction through a targeted approach to ameliorate mitochondrial dysfunction through strategies such as increasing the capacity to generate mitochondrial ATP, reducing excessive ROS overproduction, or improving the mitochondrial quality control system. Due to the significant contribution of mitochondrial dysfunction in neurodegenerative and neurovascular disorders, there is growing interest in the development of pharmacological tools to restore mitochondrial function in preclinical models of neurodegenerative and neurovascular disorders. Unfortunately, therapies that target specific components of mitochondria or oxidative stress pathways are largely unsuccessful or failed to exhibit a protective effect in clinical studies. For example, mitochondrial biogenesis plays a crucial role in the maintenance of the mitochondrial pool and activation of PPARγ increases mitochondrial biogenesis. To this end, the effect of several PPARγ agonists such as pioglitazone and rosiglitazone that increase mitochondrial biogenesis have been tested in mouse models of AD , 132, PDnditions -149. Whinditions , 151, in3.1et al. have shown that transplantation of healthy mitochondria into human breast cancer cells limits cancer cell proliferation and increases the sensitivity of the cell line to breast cancer medication, suggesting the potential of MTT for cancer therapeutics [in vitro approach, they showed that donor mitochondria could enter various cell types and improve impaired mitochondrial function. Using a rodent model of Schizophrenia, they further showed that intra-prefrontal cortex transplantation of isolated mitochondria ameliorated mitochondrial dysfunction in neurons and improved Schizophrenia-related deficit. Consistent with this study, MTT reduced brain inflammatory response and oxidative loads in the LPS-induced mouse model of depression [Mitochondrial transplantation therapy (MTT) has received increasing attention over the past few years because it presents a novel perspective of mitochondria medicine for several neural and extraneural diseases , 36, 152apeutics . Moreoveapeutics , and traapeutics , 159-162apeutics . Using apression .via axonal protrusions that were engulfed and digested by neighboring astrocytes, indicating that at least some portion of these mitochondria undergoes transcellular degradation via transmitophagy [The mechanisms by which MTT functions to increase cellular viability are not yet clear. The use of MTT dates back to 1982, when Clark and Shay first used isolated mitochondria from cells with mutations in the mitochondrial ribosomal RNA gene to confer antibiotic resistance in sensitive wild type recipient cells through coincubation . In lineitophagy . Further1). The ideal source of mitochondria may play an important role, as mitochondria can differ depending on their resident tissue. For example, the liver, heart, and skeletal muscle can exhibit slightly different mitochondrial proteomes [MTT is an innovative approach that consists of the following steps: 1) isolation of healthy and functional donor mitochondria from diverse sources, including adipose, liver, and skeletal muscle, or mesenchymal stem cells; 2) transplantation of the donor mitochondria to the host species, and finally; 3) incorporation of donor mitochondria and therapeutic benefits of MTT in host target tissues/cells  the integrity and health of the donor mitochondria, 2) how many mitochondria can be taken up by cells in a certain period, and 3) how long delivered donor mitochondria remain viable and functional following transplantation. It is well known that only functional mitochondria can confer their therapeutic efficiency. Therefore, a rapid mitochondrial isolation process and quality control check of donor mitochondria are critical for optimizing outcomes. Mitochondria can be easily and rapidly isolated both from liver and skeletal muscle tissue to allow for acute use in a clinical setting in under 30 minutes, as published by the McCully research group . This prIn vivo, mitochondrial transplantation can be carried out by several routes, including direct injection of donor mitochondria to target tissue, intranasal delivery, intra-arterial, or systemic delivery. Direct injections of autologous mitochondria in a porcine model of cardiac ischemia enhance myocardial cell viability and do not trigger inflammatory or immune responses [via tail vein injections [esponses . In a CNjections  or intrajections  or direcjections . A studyjections . While tIf the MTT is carried out through a systemic route, it is also plausible that mitochondria can distribute to several organs and elicit a possible immune response. However, several recent studies suggested that the transplanted donor mitochondria do not trigger any immune or inflammatory responses , 172. FoIn order to rapidly facilitate the transfer of donor mitochondria across the plasma membrane for increasing mitochondrial intake, a new approach was established recently, where mitochondria are conjugated with cell penetrating peptides . Mitocho3.21). Although the majority of research on mitochondrial transplantation has been reported in experimental models and in patients with cardiac injury to improve the cardiac function, there is growing evidence that mitochondrial transplantation is a feasible strategy to improve cellular functions in several preclinical models of human age-related neurodegenerative and neurovascular disorders (Table 1). However, the precise mechanisms involved in observed benefits remain unclear. Supplementing healthy mitochondria to damaged neurons promote neuronal viability, activity and neurite growth and has been shown to be effective in treating neural and extra-neural diseases [Mitochondria dysfunction is an important component of neurodegenerative and neurovascular disorders such as Alzheimer’s disease, Parkinson’s disease, and stroke. Given the critical role of mitochondria dysfunction in disease onset and progression and the therapeutic setbacks of current mitochondria-based therapy for neurodegenerative and neurovascular disorders, growing attention points to the mitochondria replacement strategy. To this end, mitochondrial transplantation therapy is an alternative approach to organelle-based therapy used in mitochondrial medicine that involves the use of healthy mitochondria to replace dysfunctional or damaged mitochondria  has approved acetylcholinesterase inhibitors galantamine, donepezil, and rivastigmine or NMDA receptor antagonist memantine, which can simply relieve the symptoms. Although the recent FDA approved the drug Aduhelm, which has been shown to directly target the amyloid beta (Aβ) pathology of AD, its efficacy is still under debate.Targeting the mitochondrial associated impairments may provide a favorable strategy due to the involvement of mitochondria in several AD-related pathologies, including oxidative stress, energy imbalance, and interaction withAβ and phosphorylated tau protein. However, several mitochondria-based therapies have been tested for murine models of AD but failed to exhibit similar neuroprotective effects in clinical studies. To overcome these limitations, MTT represents an innovative organelle-based therapy to replace dysfunctional mitochondria, thereby improving energy generation, suppressing excessive ROS production, and restoring mitochondrial function.et al. showed that systemic administration of the youthful mitochondria in aged mice reduced oxidative stress, enhanced mitochondrial function, and improved cognitive and motor function in aged mice [Supplement of healthy mitochondria into cells containing damaged mitochondria was found to be beneficial. Zhao ged mice . Consistged mice . By the ged mice . In thisged mice . Consistged mice . Thus, m3.41). For example, Chang et al. demonstrated that mitochondria conjugated with peptide infusion in the 6-hydroxydopamine (6-OHDA)-treated PC12 cells restored mitochondrial function, prevented cell toxicity, and enhanced neurite growth [Mitochondria are potential targets to slow down or reduce PD because mitochondrial dysfunction plays a central role both in sporadic and familial PD. Given the importance of mitochondrial function in PD pathogenesis, mitochondria-based therapy that replaces dysfunctional mitochondria with healthy donor mitochondria is a potentially impactful approach to counter PD. To this end, several studies examined the efficacy and feasibility of mitochondrial delivery in reducing PD progression . Nakamura et al. showed that systemic administration of placenta-derived mitochondrial reduces infarct size after cerebral ischemia-reperfusion injury in mice [via inhibition of apoptosis and reactive glia activation [Currently, the therapeutic intervention for stroke is limited owing to a restricted therapeutic time window and unsuccessful clinical translation of several neuroprotective agents. Mitochondria are an energetic hub for regulating a wide variety of cellular functions. The discovery of mitochondrial transfer from healthy astrocytes to the ischemic neurons for the purpose of ischemic injury repair and neuroprotection provides an emerging concept for designing an organelle-based therapeutic intervention in stroke . To this in mice . Interestivation . Consisttivation . Similartivation . One imptivation . Moreovein vitro approach is investigated by several labs, there has been considerable interest in testing the therapeutic benefits of MTT in experimental models of cardiac ischemia. Considering the positive outcome of MTT in cardiac ischemia, in the last few years, there has been growing interest in determining the therapeutic benefits of MTT in several experimental models of neurodegenerative and neurovascular disorders.From several decades, mitochondria have been considered to be an important therapeutic target for a variety of neurodegenerative and neurovascular disorders. Significant progress has recently been made in the development of MTT as a therapeutic strategy to improve mitochondrial function in the brain. To this end, MTT represents a novel treatment paradigm of mitochondrial transfer and replacement strategies that targets mitochondrial dysfunction in disease states. While mitochondrial transplantation between one cell to another using Replacing damaged mitochondria with mitochondria taken from healthy tissues is shaping as a promising but as yet experimental therapy to treat neurological conditions, and new breakthroughs bring MTT closer to clinical use for treating neurodegenerative and neurovascular disorders. Current data suggest that MTT significantly improves neurological complications, and at least to this point, it does not appear to produce any adverse effects in mouse models. Although early studies in humans suggest that MTT is effective in acute injuries such as cardiac ischemia reperfusion injury, yet it is also possible that MTT may be a feasible and effective treatment for other conditions, including age-related neurodegenerative disorders. Given that mitochondria are heavily implicated in the aging process, and their dysfunction results in sustained oxidative stress, metabolic dysfunction and neurodegeneration, repeated bouts of MTT may allow for the introduction of stable and healthy mitochondrial pools to substitute the dysfunctional mitochondria. In fact, MTT significantly promotes mitochondrial function, prevents neurodegeneration, and improves behavioral functions in aging and age-related neurodegenerative diseases, endorsing the application of MTT in age-related neurodegenerative diseases. However, the exact mechanism or the full therapeutic benefits of exogenous mitochondria to treat neurodegenerative and neurovascular disorders is not known, and thus, further studies are warranted. MTT is poised as a powerful tool to recapitulate a healthy metabolic environment through improvements in mitochondrial mass, ATP production, redox state, and modulation of mitochondrial biogenesis and dynamics. The finding from recent preclinical studies encourages more comprehensive and rigorous studies to examine the distribution, safety, and precise mechanisms of therapeutic efficacy of MTT which is essential for clinical translation and the expansion of therapeutic applications for several neurodegenerative and neurovascular disorders.


Saccharomyces cerevisiae cells show significant levels of cell-to-cell variability. This variability arises from de novo mutations, gene expression noise and asymmetric division. Here, I suggest that another contributing factor could be genetic drift in mitochondrial DNA (mtDNA) heteroplasmy levels. Moreover, because of the activation of general stress response pathways in cells with insufficient levels of wild type mtDNA, heteroplasmic cells can withstand certain stresses better than wild type cells. As a result, the heterogeneity within the heteroplasmic cell population could be a bet-hedging mechanism, increasing the chances of cell line survival.Individual cells within a clonal population can display pronounced phenotypic differences. In multicellular organisms, such differentiation is controlled by numerous microenvironmental, positional, and genetic cues. However, even unicellular 13 clonal somatic cells . The driving forces behind this heterogeneity include gene expression noise, cyclic processes, and asymmetry in cell divisions  The These three factors contribute to the ability of harmful mtDNA variants to persist in clonal populations along with wild type mtDNAs. Such populations should sporadically produce cells with a high proportion of mutant mtDNA and dysfunctional mitochondria. Indeed, multicellular organisms develop from a single cell, which experiences a significant genetic bottleneck in mtDNA quantity. This bottleneck decreases the diversity of mtDNA variants . DespiteEukaryotic cells are equipped with the mechanisms capable of compensating deleterious effects of mitochondrial dysfunction. In nematodes, for instance, the depletion of mitochondrial DNA triggers the expression of mitochondrial chaperones . In addiTaken together, it turns out that, on the one hand, in clonal populations mitochondrial heteroplasmy causes the sporadic occurrence of the cells with impaired mitochondrial function; and mitochondrial dysfunction activates protective mechanisms. On the other hand, the activation of such a mechanism can be useful for cells in a wide range of stressful conditions. Importantly, in a heteroplasmic cell with a high load of mutant mtDNA but retaining wild-type mtDNA the proportion of full-length wild-type mtDNA can be increased over several generations in the absence of stress. Therefore, mitochondrial heteroplasmy can be a natural way to generate phenotypic bistability in clonal populations. I suggest that bistability mediated by mitochondrial heteroplasmy can be a mechanism betting the risk in the fluctuating environment .


Emotional dysregulation symptoms following a concussion are associated with an increased risk for emotional dysregulation disorders , especially in adolescents. However, predicting the emergence or worsening of emotional dysregulation symptoms after concussion and the extent to which this predates the onset of subsequent psychiatric morbidity after injury remains challenging. Although advanced neuroimaging techniques, such as functional magnetic resonance imaging and diffusion magnetic resonance imaging, have been used to detect and monitor concussion-related brain abnormalities in research settings, their clinical utility remains limited. In this narrative review, we have performed a comprehensive search of the available literature regarding emotional regulation, adolescent concussion, and advanced neuroimaging techniques in electronic databases . We highlight clinical evidence showing the heightened susceptibility of adolescents to experiencing emotional dysregulation symptoms following a concussion. Furthermore, we describe and provide empirical support for widely used magnetic resonance imaging modalities , which are utilized to detect abnormalities in circuits responsible for emotional regulation. Additionally, we assess how these abnormalities relate to the emotional dysregulation symptoms often reported by adolescents post-injury. Yet, it remains to be determined if a progression of concussion-related abnormalities exists, especially in brain regions that undergo significant developmental changes during adolescence. We conclude that neuroimaging techniques hold potential as clinically useful tools for predicting and, ultimately, monitoring the treatment response to emotional dysregulation in adolescents following a concussion. Concussion, also known as mild traumatic brain injury (mTBI), is defined as a transitory disturbance in brain functioning due to complex pathophysiological processes induced by a traumatic event ,2. SportOn average, concussion symptoms in youth resolve within the first two weeks of injury ,8,9. HowIt is important to note that due to the rapid development of limbic regions prior to adolescence and the more gradual development of the prefrontal cortex throughout adolescence, this age period is considered a window of heightened vulnerability for psychiatric morbidity. The imbalance in maturity between limbic regions and the prefrontal cortex is thought to contribute to the typical difficulties in regulating emotions and behaviors that occur during adolescence see  19]. Th. Th19]. Emotional regulation refers to the ability to monitor, evaluate, and adapt behaviors in response to emotional stimuli . EmotionNeuroimaging techniques, including computed tomography and magnetic resonance imaging, have been widely used to objectively characterize neurodevelopmental changes as they occur throughout the lifespan ,26,27,28The goal of this narrative review is to highlight current evidence on the utility of advanced neuroimaging techniques in investigating emotional dysregulation in adolescents following concussions. To this end, we provide a comprehensive narrative review of the available literature in electronic databases  using keywords such as concussion, post-concussion symptoms, emotional dysregulation, psychological symptoms, psychiatric symptoms, mood symptoms, depression, anxiety, neuroimaging, functional magnetic resonance imaging, and diffusion magnetic resonance imaging. An additional search was conducted using keywords such as children, adolescent, adolescence, youth, and adult to identify studies reporting findings in different age ranges between 2010 and 2023. To provide the reader with a conceptual framework, we will (1) review emotional dysregulation following concussion from a neurodevelopmental perspective; (2) provide an overview of how the employment of different neuroimaging modalities could enhance our understanding of the neuropathophysiological mechanisms driving emotional dysregulation in adolescent concussion; and (3) highlight neuroimaging findings linking concussion and emotional dysregulation from the existing literature. With this work, we seek to convey the breadth and depth of available research on the utility of advanced neuroimaging techniques in investigating emotional dysregulation in adolescent concussions. Additionally, we aim to identify research gaps and provide recommendations for future research in this area.N = 14,765) sample of high school students in the United States, 36% of students with a concussion history reported persistent depressive symptoms for up to 12 months . Th. Th52]. Within and between these systems, the subregions of the prefrontal cortex are directly and indirectly connected to other cortical and subcortical regions by major white matter tracts . The infDuring adolescence, prefrontal cortical regions undergo significant structural and functional changes . As adolConcussions can damage the brain by either direct collision or the acceleration/deceleration forces associated with impact . Direct Neuroimaging procedures offer a way to examine the pathophysiologic processes of concussion in vivo. Quick and capable of detecting a broad spectrum of pathologies, computed tomography is the current standard neuroimaging procedure for brain injuries on the day of injury. However, patients with concussion typically undergo neither computed tomography nor ‘conventional’ magnetic resonance imaging as the diagnostic capacity of these procedures for mild forms of brain injury is limited . In factOver the past fifteen years, the employment of advanced magnetic resonance imaging methods has promoted a better understanding of the pathophysiologic mechanisms of concussion. Their use in longitudinal designs is likely to shed light on important mechanisms underpinning the acute, sub-acute, and chronic stages of concussion. For example, in adolescence, these approaches are well-suited to determine if a progression of concussion-related abnormalities exists in chronic stages of concussion after disentangling the contribution of the neurodevelopmental changes typical of these ages.Broadly, functional magnetic resonance imaging measures the hemodynamic response, the Blood-oxygen-level-dependent (BOLD) signal, as a proxy of neural activity. This activity corresponds to the oxygen consumption of firing neurons, and it can be assessed either during rest (resting state magnetic resonance imaging) or during task performance (task magnetic resonance imaging) ,91. In aWhen analyzing resting-state magnetic resonance imaging, researchers generally focus on functional segregation or functional integration across networks ,94,95,96Task magnetic resonance imaging maps the response of the brain to tasks administered during imaging collection . DiffereOn the other hand, diffusion magnetic resonance imaging provides indices of tissue microarchitecture by measuring the diffusion imaging properties of the water molecules in the brain tissue ,121,122.Advances in diffusion magnetic resonance imaging have increased our ability to characterize micro- and macro-structural properties of different brain tissues and detect abnormalities across different medical conditions  ,124. StaStudies using diffusion magnetic resonance imaging have found concussion-related microstructural abnormalities in several brain regions ,139,140,While functional and diffusion imaging have been mostly used to document brain abnormalities in concussion, other modalities  have also been used.Susceptibility-weighted imaging is susceptible to the parametric properties of different blood products of micro- and macro-hemorrhage . In concussion, employing this technique allows the detection of micro-hemorrhagic lesions that would otherwise go unnoticed  . Some stMagnetic resonance spectroscopy is a metabolic imaging method that detects and quantifies metabolites  in the brain tissue . In concThere has been a paucity of studies examining the structural and functional neural changes accompanying emotional dysregulation in adolescent concussion, especially as to whether detecting concussion-related abnormalities in the brain relates to psychiatric vulnerability following concussion . ImportaAnimal studies have shown increased anxiety-related behaviors after a concussion, as measured by the locomotor activity of concussed rats in a maze. These behaviors were associated with increased neuronal excitability of the amygdala, as demonstrated by decreased inhibitory synaptic transmission . AnotherIn the human literature, children and adolescents appear to have abnormal neural responses to emotional stimuli after injury ,184. SpeEvidence from adults suggests that white matter abnormalities in those who developed depressive symptoms following concussion do not differ from those found in adults with depression alone . Specifin = 15) who developed full or subclinical forms of depression or anxiety six months post-injury. Most of them (60%) had no prior psychiatric history [To our knowledge, few studies have examined the relationship between the concussed brain and the emergence of depression or anxiety in children and adolescents. Max, Keatley  found ev history . Our rec history . Compare history  in the c history . These w history . A lower history . These fAs noted above, adolescence is a sensitive developmental window characterized by structural and functional changes reflecting brain plasticity, and injury may induce a risk for psychiatric morbidity. A key aspect that needs further investigation is whether a concussion in the adolescent brain can affect the establishment of connections that support emotion regulation. This may lead to heightened vulnerability to anxiety and depression later in life. As noted above, adolescence is a sensitive developmental window characterized by structural and functional changes reflecting brain plasticity, and injury may induce risk. However, the lack of longitudinal studies, the paucity of studies in youth, and heterogeneity due to a lack of standardized protocols across studies contribute to this knowledge gap. These factors limit the interpretability of the existing findings and their clinical and translational relevance .To properly address this question, longitudinal neuroimaging studies are needed to disentangle the effects of concussion on the adolescent brain from those associated with typical development or those related to preexisting psychiatric vulnerability  and morbidity . Altogether, the literature reviewed in this section provides support for the employment of advanced neuroimaging protocols as a tool to determine the effect of concussions on the developing brain. Moreover, it further determines if a progression of concussion-related lesions in brain regions involved in emotional regulation relates to a heightened vulnerability for psychiatric morbidity in adolescents who sustain a concussion. However, the lack of longitudinal studies, the paucity of studies in youth, and heterogeneity due to a lack of standardized protocols across studies limit the interpretability of these findings and their clinical translational relevance .Understanding the neural mechanisms of emotional dysregulation following concussion in adolescents may promote early identification of at-risk adolescents. Specifically, based on the networks structurally and/or functionally affected by concussion, advanced neuroimaging techniques may help (1) assess risk for persistent emotional dysregulation symptoms and psychiatric morbidity, (2) tailor individualized treatment approaches , (3) track recovery progress, and (4) inform decisions regarding return to activities. The identification of specific regions implicated in emotional dysregulation following concussion in adolescents might also pave the way for the development and inclusion of novel personalized treatments in clinical practice. One such example is neurofeedback, directed at modulating the functioning of identified targeted regions that may be key for recovery in adolescent concussion. Neurofeedback is a form of biofeedback technique that aims to train individuals to consciously control their brain activity . CurrentThere is a paucity of studies directly examining the relationship between concussion-related brain abnormalities and heightened vulnerability to emotional dysregulation in adolescents. The identification of structural and/or functional abnormalities in emotional regulation circuits may serve as biomarkers of emotional dysregulation in adolescent concussion and promote the identification of those adolescents who are most at risk for psychiatric morbidity following concussion. To achieve this, future studies should employ advanced neuroimaging techniques in longitudinal designs that allow for the identification of functional and structural changes in regions and networks involved with emotional regulation and the effects of environmental factors  on brain recovery post-concussion. However, there are important steps to consider before contemplating their implementation into clinical practice: (1) Standardize neuroimaging protocols and analytic methods to ensure consistency and facilitate comparison across different sites; (2) establish data sharing protocols to promote replication of findings incorporating different socio–demographic factors ; (3) foster communication between researchers and clinical providers to facilitate the translational aspect of concussion neuroimaging research; and (4) perform validation studies that address the clinical relevance and cost-effectiveness of these techniques in clinical settings.Although variability in study design, age ranges, sample sizes, and post-injury time points limits conclusions, the literature reviewed above supports the employment of advanced magnetic resonance imaging techniques in adolescent concussions. These techniques have the potential to achieve several key objectives: (1) promote a better understanding of the different pathophysiologic mechanisms of acute, subacute, and chronic stages of concussion; (2) discern between structural and functional abnormalities associated with a concussion from those associated with normative development or preexisting psychopathology, which may overlap (3) foster early identification of risk for psychiatric outcomes; and (4) serve as a prognosis tool for returning to activities. These can be instrumental in improving clinical decision-making and fostering the identification of early intervention strategies aimed at strengthening emotional and behavioral regulation strategies in at-risk groups.


Syndromic Logger) in the emergency room of a large hospital. The platform captured syndromic signals from audio, thermal imaging, and radar, while the ground truth data were collected from the hospital’s electronic health record. Our analysis revealed a significant correlation between the aggregated cough count and positive COVID-19 cases in the hospital . Notably, this correlation was higher than that observed with the number of individuals presenting with fever , a widely used syndromic signal and screening tool for such diseases. Furthermore, we demonstrate how the data obtained from our Syndromic Logger platform could be leveraged to estimate various COVID-19-related statistics using multiple modeling approaches. Our findings highlight the efficacy of aggregated cough count as a valuable syndromic indicator associated with the occurrence of COVID-19 cases. Incorporating this signal into syndromic surveillance systems for such diseases can significantly enhance overall resilience against future public health challenges, such as emerging disease outbreaks or pandemics.Syndromic surveillance is an effective tool for enabling the timely detection of infectious disease outbreaks and facilitating the implementation of effective mitigation strategies by public health authorities. While various information sources are currently utilized to collect syndromic signal data for analysis, the aggregated measurement of cough, an important symptom for many illnesses, is not widely employed as a syndromic signal. With recent advancements in ubiquitous sensing technologies, it becomes feasible to continuously measure population-level cough incidence in a contactless, unobtrusive, and automated manner. In this work, we demonstrate the utility of monitoring aggregated cough count as a syndromic indicator to estimate COVID-19 cases. In our study, we deployed a sensor-based platform ( The act of coughing is part of the body’s defense system that uses automatic reflex mechanisms to clear airways of foreign substances, irritants and excessive mucus. Cough and related complaints have been recognized as a cardinal symptom or syndromic signal of respiratory illnesses, such as the SARS-CoV-2, influenza and influenza-like illness, asthma, chronic obstructive pulmonary disease (COPD), and lung cancer. Recent progress with cough assessment has led to improved diagnosis and more effective disease management for individual patients , 18. HowSyndromic Logger is the next-generation contactless sensing platform..https://Based on our findings, we propose the integration of a sensor-based, contactless, and unobtrusive platform as a valuable augmentation to existing approaches, such as body temperature-based screening, in diverse public environments. Our results demonstrate that sensor-based platforms have the ability to capture clinically relevant syndromic signals, specifically the aggregated cough count. This capability allows us to overcome the limitations of existing methods which include reliance on manual human logging and reporting, as well as associated reporting delays. By embracing the adoption of sensor-based platforms, we can enhance the effectiveness and efficiency of public health surveillance, providing a superior solution for detecting and monitoring public health situations to overcome potential challenges in the future.


Bronchial and alveolar remodeling and impaired epithelial function are characteristics of chronic respiratory diseases. In these patients, an increased number of mast cells (MCs) positive for serine proteases, tryptase and chymase, infiltrate the epithelium and alveolar parenchyma. However, little is known regarding the implication of intraepithelial MCs on the local environment, such as epithelial cell function and properties. In this study, we investigated whether MC tryptase is involved in bronchial and alveolar remodeling and the mechanisms of regulation during inflammation. Using novel holographic live cell imaging, we found that MC tryptase enhanced human bronchial and alveolar epithelial cell growth and shortened the cell division intervals. The elevated cell growth induced by tryptase remained in a pro-inflammatory state. Tryptase also increased the expression of the anti-apoptotic protein BIRC3, as well as growth factor release in epithelial cells. Thus, our data imply that the intraepithelial and alveolar MC release of tryptase may play a critical role in disturbing bronchial epithelial and alveolar homeostasis by altering cell growth–death regulation. Human mast cells (MCs) are immune cells strategically located at sites close to the external environment, such as the skin, gut and lungs, that play a crucial role in the first-line innate defense against bacteria, viruses and other foreign pathogens . Upon acAirway and epithelial remodeling are pathological characteristics of chronic respiratory diseases, and involve alterations in the composition of the airway architecture in both the large and small airways as well as of the alveolar parenchyma. Eventually, these events lead to impaired airflow and ventilation. Although MCs are speculated to have an emerging role in disease progression ,13, therThe bronchial and alveolar epithelium are under constant renewal and require tight regulation of cell growth, apoptosis and necrosis to maintain proper and fully functional barrier properties. Cell death signaling pathways can be activated upon stimuli such as infection and damage, and are important in regulating homeostasis within the airway epithelium. BIRC3 is a central anti-apoptotic protein and controls cell death homeostasis by regulating important signaling events in the caspase cascade . EpithelWe have previously shown that bronchial epithelial cells treated with tryptase upregulate the expression of the proliferation marker Ki67 and elevate mitochondrial activity. This was associated with enhanced proliferation; however, there was no effect on cell growth in tryptase-treated cells . Althoug2 until cell confluency reached 80–90%. All experiments on BECs and AECs were performed in starvation medium containing RPMI or F12 medium with 1% FBS and 1% penicillin–streptomycin. Primary human BECs were collected from patients with mild to moderate asthma who were not treated with inhaled corticosteroids for at least 3 months and had airway hyperresponsiveness to mannitol. Patients underwent bronchoscopy with endobronchial brush biopsies as part of the RECONSTRUCT study  thymidine incorporation, which are indications of proliferative effects, migration and increased gap closure in BECs ,32,33, bTo investigate whether the cell growth alterations, which were found in the cell lines, were mediated via PAR2, inhibitor I-191 was used. Although a small dampening in cell growth could be observed with I-191, the effect was not significant. This might indicate that PAR-2 signaling is less important for cell growth in AECs compared to BECs, where the effect is more pronounced . In a reFurthermore, we wanted to investigate whether the cell division pattern was altered by tryptase. Interestingly, we found that tryptase-treated BECs had a shorter cell division interval in the later phase of the experiment, i.e., after the first and second cell division, whereas in alveolar epithelial cells, tryptase had a faster action and had induced a shorter cell division rate already at an earlier time point. Interestingly, these changes persisted up to 72 h. We have previously shown that the majority of the enzymatic activity of tryptase is lost after approximately 6–12 h post-stimulation . We haveTCs infiltrate the alveolar parenchyma in many respiratory diseases, which is also strongly affected by remodeling, we aimed to investigate the tryptase effect of GF release in alveolar epithelial cells. In the present study, we demonstrated, for the first time, that tryptase significantly increases GF release in both primary BECs from patients with asthma and also in AECs. Luminex data from tryptase-treated primary cells revealed that tryptase shifted the GF profile since it upregulated several GFs  and decreased others  when compared to non-stimulated cells. These findings again emphasize the detrimental role of MCs in chronic respiratory disease pathogenesis, since these GFs have strong cell growth and chemotactic effects on multiple structural cell types [A broad range of cytokines and GFs, e.g., basic FGF and PDGF-AA, act as central factors in epithelial and alveolar remodeling and respiratory disease progression due to their potent mitogenic and chemoattractive properties. Although GFs are crucial for tissue maintenance and survival, an overexpression or dysregulation of GFs have detrimental roles and are commonly seen in chronic respiratory diseases, including asthma, COPD and IPF. The underpinning mechanisms of altered GF homeostasis in alveolar parenchyma and airways are not fully understood ,38,39,40ll types ,37,41,42The cellular inhibitor of apoptosis proteins (cIAP)2, also known as BIRC3, is an anti-apoptotic protein, which is highly important in the control of cell death–survival regulation. In recent years, it has particularly been highlighted for its contribution in cancer progression by inhibiting apoptosis and thus promoting uncontrolled cancer cell proliferation and tumor survival. BIRC3 exerts pro-survival functions by regulating caspases and pro-caspases ,43,44. DSince patients with asthma show augmented chronic airway inflammation and viral-induced exacerbations, we further wanted to study the consequence of tryptase on airway cell growth in pro-inflammatory and viral environments. We used three agents strongly associated with the induction of airway inflammation and viral responses: the viral mimic polyI:C, LPS and cigarette smoke extract (CSE). Our results showed that the elevation in BEC growth induced by tryptase remained in a pro-inflammatory environment (LPS and CSE). PolyI:C-stimulated cells showed a tendency toward poorer cell growth that, unlike LPS and CSE, did not increase in the presence of tryptase. Interestingly, viral infections are the main cause of exacerbations and epithelial damage in asthmatic patients, which also show impaired healing capacity in the bronchial epithelium ,47. HoweIn this study, our results reveal that tryptase exerts the ability to induce cell growth and the shortening of cell division intervals, and alter anti-apoptotic regulation and growth factor release in both human alveolar and bronchial epithelial cells. In addition, we show that tryptase-treated primary bronchial epithelial cells from asthma patients enhanced the protein expression of the anti-apoptotic protein BIRC3, altered growth factor release and elevated the expression of the proliferative marker Ki67, although no functional difference was observed. Taken together, our findings imply that intraepithelial MCs and their release of tryptase may be key players in airway remodeling by disturbing normal cell growth–death homeostasis and hence promote epithelial cell growth and survival and possibly other structural cells in the lung tissue as well. Apart from advancing the concept of the role of MCs in chronic respiratory diseases, these observations may provide important insights into how to improve treatment strategies by targeting tryptase.