Platelets as Hidden Drivers of Hepatitis C Pathogenesis

1. Overview

(1) Scientific Rationale and Importance

Hepatitis C virus (HCV) infection remains a major global health burden, particularly in the Middle East and North Africa (MENA) region. While HCV pathogenesis has traditionally been studied in the context of hepatocytes and immune cells, increasing evidence suggests that platelets play a critical role beyond hemostasis, functioning as immune-modulatory and pro-inflammatory cells.

Our preliminary data demonstrate that:

Platelets from HCV patients display hyperactivation and altered cytokine profiles.
HCV RNA can be detected in a subset of circulating platelets.
Platelet inflammatory and procoagulant phenotypes differ between early and advanced disease stages.

We hypothesize that platelets contribute to:

Viral dissemination,
Immune dysregulation,
Thrombo-inflammatory complications associated with chronic HCV.

By integrating high-dimensional immune profiling, platelet functional imaging, and receptor-binding analyses, this project will define the molecular mechanisms underlying platelet–HCV interactions. The findings may reveal novel biomarkers of disease progression and identify platelet-specific therapeutic targets aimed at reducing inflammation and thrombotic risk in HCV patients.

This work is particularly important in the MENA region, where HCV prevalence remains high and where understanding host–virus interactions could improve stratification and management strategies.

(2) Confirmation of Funding, Facilities, and Ethics Approvals

Facilities:
All experimental procedures will be conducted in fully equipped laboratories at:

Mohammed V University (Rabat, Morocco),
Qatar University (Doha, Qatar),
Collaborating Canadian institutions (Montreal Heart Institute, University of Calgary).

Available infrastructure includes:

Flow cytometry and mass cytometry platforms,
Confocal and high-resolution platelet imaging systems,
Molecular biology and RT-PCR facilities,
Radioligand binding and protein analysis platforms,
Access to clinical recruitment sites (Cheikh Zaïd Hospital, HMC).

Ethics Approvals:
The study has obtained Institutional Review Board (IRB) approval from Cheikh Zaïd Hospital in Rabat and complies with the Declaration of Helsinki. Additional approvals at participating institutions are in place or will be maintained for sample transfer and data sharing in accordance with institutional and national regulations.

Funding and Institutional Support:
Core laboratory infrastructure and clinical recruitment pipelines are supported by institutional resources at participating universities. The project leverages established collaborations and existing patient cohorts (200+ recruited patients), ensuring feasibility and cost-efficiency.

(3) Estimated Timeline

The project is designed over a 24-month period:

Months 1–6

Final regulatory confirmations
Continued patient recruitment
Standardization of platelet–HCV interaction assays

Months 6–18

High-dimensional platelet phenotyping
TSPO expression and binding studies
Receptor identification and validation experiments
Functional inhibition assays

Months 18–24

Statistical modeling and correlation with clinical parameters
Integration of multi-omics datasets
Manuscript preparation and dissemination of findings

Parallel experimentation and recruitment ensure efficient use of time, and much of the preliminary groundwork (cohort recruitment, assay optimization) has already been initiated.

(4) Data Sharing Commitment

We are committed to transparent and responsible data sharing in accordance with international open science standards.

De-identified clinical and experimental datasets will be made available upon publication through institutional repositories or recognized public platforms.
Flow cytometry and mass cytometry datasets will be deposited in appropriate public repositories.
Protocols and analytical workflows will be shared to ensure reproducibility.
Publications will be submitted to peer-reviewed journals with open-access options whenever feasible.

All data sharing will strictly comply with patient confidentiality regulations and institutional data governance policies.

2. Introduction

Platelets are traditionally recognized for their role in hemostasis and thrombosis. However, over the past two decades, they have emerged as central regulators of inflammation and innate immunity. Platelets express multiple immune receptors, including Toll-like receptors (TLRs), Fc receptors, C-type lectin receptors, and immunoreceptor tyrosine-based activation motif (ITAM) receptors. They can sense pathogens, release inflammatory mediators, interact with leukocytes, and contribute to thrombo-inflammatory responses.

In viral infections, platelets have been shown to:

Internalize viral RNA,
Become hyperactivated,
Release cytokines and chemokines,
Facilitate immune cell recruitment,
Contribute to microvascular thrombosis.

Our previous work demonstrated that platelets can associate with viral RNA and become hyperactivated during SARS-CoV-2 infection (Zaid et al., Circ Res 2020), supporting the concept that platelets may act as immune sentinels and viral carriers.

Hepatitis C virus (HCV) infection remains highly prevalent in the MENA region and is associated not only with liver disease but also with systemic inflammation and cardiovascular complications. While hepatocytes are the primary site of infection, extrahepatic manifestations suggest broader host–virus interactions. Emerging evidence indicates that:

Viral RNA may be detected in circulating platelets,
Chronic HCV infection is associated with inflammatory cytokine dysregulation,
Thrombotic risk is increased in certain HCV patients.

However, the molecular mechanisms by which platelets interact with HCV, and whether they actively contribute to viral dissemination and thrombo-inflammatory signaling, remain poorly defined.

This project builds on the theoretical framework that platelets function at the interface of immunity, inflammation, and thrombosis, and may serve as active participants in HCV pathogenesis rather than passive bystanders.

Research Questions

Do circulating platelets directly interact with HCV in infected patients?
Does HCV alter platelet activation state, secretory profile, and procoagulant membrane dynamics?
Are specific platelet receptors responsible for HCV binding and signaling?
Is platelet TSPO expression modulated during HCV infection, and does it correlate with inflammatory phenotype or disease stage?
Do platelet phenotypes correlate with clinical severity (early vs advanced HCV)?

Hypotheses

H1: Platelets from HCV-infected patients contain detectable viral RNA and/or viral proteins, indicating direct platelet–virus association.

H2: HCV infection induces a hyperactivated and procoagulant platelet phenotype characterized by increased P-selectin expression, phosphatidylserine exposure, cytokine release, and enhanced prothrombinase activity.

H3: Specific platelet surface or mitochondrial receptors (e.g., TSPO, Fc receptors, C-type lectin receptors) mediate HCV binding and contribute to downstream activation signaling.

H4: Platelet TSPO expression and ligand-binding capacity are altered during active HCV infection and correlate with inflammatory cytokine levels and disease severity.

H5: Distinct platelet immunophenotypic signatures differentiate early-stage from advanced-stage HCV and may serve as biomarkers of progression.

Impact Statement

Regardless of whether our hypotheses are fully confirmed or partially refuted, this study will significantly advance understanding of platelet biology in chronic viral infection.

If our hypotheses are confirmed, the findings would:

Establish platelets as active contributors to HCV pathogenesis,
Reveal novel host–virus interaction mechanisms,
Identify platelet-specific biomarkers of disease progression,
Provide rationale for targeting platelet signaling pathways as adjunctive therapeutic strategies.

If our hypotheses are not confirmed, the study will still:

Clarify the limits of platelet involvement in HCV infection,
Refine current models of thrombo-inflammatory signaling in chronic viral disease,
Generate high-dimensional platelet datasets valuable for future mechanistic studies.

In both scenarios, the project strengthens the conceptual framework linking immunity, thrombosis, and viral infection, and may have broader implications for other chronic viral diseases beyond HCV.

3. Methods

Participants/Sample

Consecutive patients with confirmed HCV (total of 200 patients) including early and advanced forms admitted at Cheikh Zaïd Hospital from October 17, 2023 to present. The male/female ratio was 1.19:1. The mean age of patients was 54.37 ± 18.15 with 56 ± 16.28 and 51 ±21.41 years for females and males respectively. Control group for plasma analysis assays and experiments on platelets lysates and aggregation is selected to be sex and age matched with HCV patients with a male/female ratio of 1.22:1 and a mean age of 50.75 ± 17.88 years. The overall study design is not blinded, as patients are initially classified by symptoms, presence of viral RNA, and disease severity. These parameters are known when clinicians collect blood. However, the analyses are blinded in the following assays: measurements of cytokine profiles, extracellular-vesicle measurements, PF4, and serotonin.

Materials and Procedures

AIM 1. Perform high-resolution multidimensional characterization of platelets in HCV patients.

Preliminary data. Human blood was collected from individuals suspected of Hepatitis C infection (proximity to infected patients, presence of symptoms) at Cheikh Zaïd Hospital in Rabat (Morocco) by Co-LPI Dr Y. Zaid, and platelets were freshly isolated. RT-PCR was used to determine HCV infection. Cytokine levels in plasma and platelets were assessed using Bio-Plex Pro Human Cytokine 27-plex Assay (Bio-Rad, Marnes-la-Coquette, France). If cytokines were not detected, a concentration corresponding to the lowest value extrapolated from the standard curve was attributed to the sample for quantification and comparison analyses. We made the seminal finding that the concentrations of eight cytokines or growth factors were significantly increased (G-CSF, IFN-γ, IL-1β, IP-10, MIP-1β, MDC, RANTES, TNFα, VEGF-A); the concentrations of two cytokines (IL-9 and IL-15) were significantly decreased, while that of 17 cytokines or growth-factors were similar to healthy controls (Fig.1). Elevated inflammatory cytokine levels are reported in the blood of patients with CD and may contribute to the overwhelming inflammation.

Figure 1. Heat map visualization of 27 cytokine expression profiles in plasma and platelet content of patients with Hepatitis C in early stage (median of n = 20) versus advanced stage (n = 16). Cytokine expression is represented as a (log2) fold change relative to healthy donors. The numbers in each part represent the change and statistical significance (in brackets), and the color codes refer to brown for increased expression and blue for decreased expression.

Experimental design. Heparinized blood from HCV early stage (no cirrhosis) and advanced stage (cirrhosis) will be collected at Cheikh Zaïd Hospital by Dr Y. Zaid's group. For all patients, sex, age, prior medication and condition, duration of hospitalization or mortality, treatment, and complete blood count will be documented. Patients with chronic diseases and those who took drugs that interfere with platelet function within 2 weeks prior to experimentation are excluded from this study. Cells will be labeled with metal-conjugated antibodies, fixed per manufacturer instructions, and sent to the flow cytometry platform at CHU de Quebec for batch analyses. Data will be analyzed using the Astrolab workflow. Data from males and females will be processed and presented separately. Mass cytometry characterization of platelets in HCV patients. In order to determine potential mechanisms promoting platelet function during Hepatitis C, the complete set of platelet receptors involved in thrombosis and inflammation will be evaluated using mass cytometry.

Metal conjugated antibodies against full set of platelet activation-regulatory receptors, including Immunoreceptor tyrosine-based activation motif (ITAM) receptors (glycoprotein VI, c-type lectin 2 and FcgRIIA) (16, 17) and regulatory immunoreceptor tyrosine-based inhibition motif (ITIM)-containing receptors (G6b-B, PECAM-1 and leukocyte-associated immunoglobulin-like receptor)(34) will be used to fully characterize surface receptors. Moreover, the IgG, IgA, and IgE platelet receptors (FcyRIIA, FcaRI, FcεRI) will also be examined to determine their potential role in platelet-virus interactions.

Immune profiling assay of platelet-immune cell interactions. As platelet-leukocyte interactions are reported in inflammatory diseases and contribute to leukocyte extravasation or organ invasion (7, 14), the interactions of platelets with leukocytes will be evaluated using mass cytometry. For this, we will utilize the Maxpar® Direct™ Immune Profiling Assay™ (Fluidigm), to which metal-conjugated anti-CD41 antibody will be included, thus permitting the complete assessment of 37 different immune cell populations in blood and the occurrence of the interaction of these cells with platelets.

1.3 Platelet secretome characterization during HCV infection. As for now, fixed platelets, platelet lysates, and plasma (platelet-free plasma) have been collected from 42 healthy individuals (hospital personnel confirmed HCV negative), and 200 patients with Hepatitis C, early and advanced stages (Fig.1). Plasma and platelet lysates will be examined for their content in PF4 and serotonin (ELISA tests). This approach will reveal whether alpha (PF4) and dense granules (serotonin) content were released. Quantification of various subtypes of platelet CD41+ microparticles (exposing or not phosphatidylserine, containing or not mitochondria) will also be performed as we reported (15).

1.4 Identification of prevalent IgG, IgA, and IgE recognizing HCV In all plasma samples, antibodies (IgG, IgA, and IgE) will be searched using serology assays. These observations will enable us to deduce associations between the presence of antibodies (pre-existing and specific to HCV) and platelet activation and/or cellular interactions through specific receptors.

1.5 Detailed fluorescent imaging to characterize platelet procoagulant membrane dynamics (PMD) in HCV patients. We will utilize high-resolution 3D confocal imaging coupled with the systematic analysis of PMD. In vivo, platelets adhere to solid matrixes such as the vessel wall, where unique morphological transformations integral to platelet prothrombotic responses can occur (9,10). These processes cannot be readily assessed by conventional in vitro platelet function tests, which rely on surrogate measures of platelet ability to aggregate, such as aggregometry assays. The systematic analysis of platelet PMD offers a solution by using a high-resolution 4D imaging approach to directly examine platelet-blood cell interactions and the spatiotemporal dynamics of the morphological and functional changes platelets undergo (Fig.3)(9,10,12,13). Platelet PMD is increasingly being studied in translational research23–25, as it provides high-resolution elucidation of the cell biology of thrombosis and hemostasis. We will assess platelet adhesion and procoagulant morphological changes over inert surfaces (as in Fig.2) and matrixes coated with collagen (type I, IV), fibrinogen, or VWF under static and flow conditions. The formation of thrombin on the platelet membranes will be visualized using the Alexa-Fluor 488/647 conjugated thrombin antibody. The antibody is specific for an epitope mapping between amino acids 331-376 within an internal region of thrombin of human origin. Parallel experiments will use the fluorogenic thrombin substrate Z-Gly-Gly-Arg aminomethyl coumarin (ZGGR-AMC), as shown (Fig.2ii-iii). We will probe for the assembly of coagulation factors of the prothrombinase complex on the platelet membrane, as we have previously done(18, 19, 20). We will also assess the generation of procoagulant MVs using flow cytometric and imaging approaches as described(21). Agonist-induced formation of distinct platelet phenotypes in HCV patients will be assessed (Aim 1) using our established imaging approach(18, 19, 20, 22). We will utilize Volocity® software for data deconvolution, analysis, 3D/4D re-construction, and video rendition, as previously done(18, 19, 20, 22).

Figure 2. Platelet PMD Imaging (i): 3D reconstruction of collagen-activated platelets showing ballooned and procoagulant platelets (red-annexin-v) surrounded by conventionally spread non-ballooned platelets17. (ii): 3D reconstruction of fluorogenic thrombin substrate (cyan) on the ballooned platelets and microvesicles membrane. (iii): time course of thrombin generation in a representative single platelet after adhesion to the collagen-coated surface. (iv): α-granule mobilization of P-selectin (blue) to the ballooned and spread membrane of procoagulant platelets (v): aggregating platelets over exposed collagen. Yellow arrows indicate the direction of aggregation and procoagulant nidus.

1.5.2 Granule secretion in HCV platelets will be investigated by full concentration-response measurements and kinetic assessments through time courses. Secretion of dense granules will be assessed luminometrically, and granule numbers will be determined by transmission electron microscopy (TEM). α-granule secretion will be assessed by imaging and flow cytometry determination of P-selectin surface expression and ELISA assessment of TGFβ and PF4. Granules will be visualized by staining for 5HT or P-selectin to determine if granule centralization (a key feature of the secretory process) is disrupted or augmented in platelets of patients with HCV infection.

1.5.3 Surface receptors expression: We will use high-resolution imaging and flow cytometry to assess the expression of surface receptors, including PECAM-1/CD31, CD41, GPVI, GPIX, α2β1 integrin, GPIbα, and αIIbβ3, and follow their co-localization with plasma anti-β2GPI, C5a, or C5b-9. We will probe for the high-affinity form of αIIbβ3 as an extension of the assessment of activation.

Overall, our approach to Aim-1 will fully characterize the platelet role in HCV patients during infection and further demonstrate the interaction of other blood cells with platelets. In addition, our approach will indicate what receptors and kinases are relevant to platelet aggregation, antibody response, or virus interactions that are expressed during HCV infection. The study will be the first to define the platelet secretome and the presence of extracellular vesicles, such as those containing mitochondrial damage-associated molecular patterns in HCV patients (23, 24). Additionally, by characterizing platelet procoagulant potential in patients with HCV infection, we will further understand the prothrombotic morphological changes platelets undergo in these patients. This, in turn, may inform antithrombosis regimen practice in the management of patients with HCV infection.

AIM 2. Characterize the expression, localization, and binding characteristics of the 18 kDa translocator protein (TSPO) in various stages of HCV infection.

Preliminary data. We identified HCV RNA in platelets of more than 12% of the patients analyzed (4/27 HCV patients tested, none in healthy subjects), suggesting that HCV may infect platelets (or megakaryocytes) and may be disseminated through the whole organism using platelets as a vehicle.

2.1 Identification of HCV viral proteins in platelets of HCV patients. While HCV RNA was identified in platelets in 12% of the HCV-positive patients tested, we will confirm the presence of HCV in all available samples (n=200, 109 males and 91 females). RT-PCR will be used to detect the presence of the HCV genome. With this approach, we may reveal associations between the presence in platelets of HCV RNA and clinical outcomes (e.g., +/-cirrhosis, survival, or death). For all experiments, data obtained using specimens from males and females will be presented separately to determine whether sex is a variable in our analyses.

Platelets will be prepared from the platelet-rich plasma, and the platelet membranes will be homogenized and centrifuged for 30 minutes at 49,000g. The pellet will be suspended in PBS and used for [3H]PK 11195 binding to determine the affinity and density of platelet TSPO, and a Western blot analysis will be conducted. We will determine TSPO binding and protein expression in the various stages of the HCV infection. This will be the first study of the impact of HCV infection on TSPO expression. Since TSPO was found by our group to exhibit anti-inflammatory activity (25, 26, 27), we assume that in the active stages of the HCV infection, there will be an elevation in platelet TSPO density and expression in an attempt to counteract the inflammation, while recovery or clinical improvement will be associated with a gradual decline in TSPO expression in correlation with a parallel decrease in the pro-inflammatory cytokines.

Platelet TSPO may play a role in infection-induced platelet signaling and hyperreactivity. Since TSPO is a mitochondrial transporter that was found by our group to exhibit anti-inflammatory activity. Including suppression of proinflammatory cytokines and chemokine release and NF-κB κB expression in macrophage and microglial cells stimulated by LPS. In a cellular model of HCV, the TSPO ligand etifoxine administration increased the TSPO protein expression and recovered the HCV-mediated lower mitochondrial membrane potential (MMP) without affecting HCV infection. Etifoxine also reversed the HCV-induced lipid accumulation by modulating the expressions of sterol regulatory element-binding protein-1 and apolipoprotein (28). An analysis of the gene expression profile demonstrated that TSPO interaction with PPIB might play an important role in liver damage in alcoholic hepatitis (29). We assume that in the active stages of the HCV infection, there will be an elevation in platelet TSPO density and expression in an attempt to counteract the inflammation, while recovery or clinical improvement will be associated with a gradual decline in TSPO expression in correlation with a parallel decrease in pro-inflammatory cytokines and chemokines.

Expected results: While we are confident that HCV RNA is transported by platelets, this aim will confirm the presence of actual viral particles in platelets, which may be associated with certain clinical outcomes. If platelets are hyperactivated and present in a given phenotype (determined in AIM1), this may indicate that the virus in platelets mediates the activation.

Pitfalls. Sufficient statistical power is required to perform association analyses. At the moment, we already have 200 patients recruited for whom we have isolated platelets. However, by the end of the project, we anticipate that 400 samples will be collected. As approximately 12% of the patients show HCV RNA in their platelets, statistical power will be sufficient. Analyses will be performed by a biostatistician.

AIM 3. Identify candidate receptors expressed by platelets implicated in HCV interactions.

Our approach will reveal the expression pattern of candidate receptors capable of mediating interactions (directly or indirectly through immunization) with HCV. The expression may vary with comorbidities, which may explain how the virus can impact different individuals differently as a function of pre-existing liver disease. Moreover, by adding the MK markers to our immunofluorescence studies, we will further determine how MK may interact with HCV.

Planned Analyses

Analysis Pipeline

All datasets (clinical, RT-PCR, flow cytometry, cytokines, imaging, TSPO binding, receptor assays) will be merged using unique patient identifiers. Analyses will be conducted in three groups: early-stage HCV, advanced-stage HCV, and healthy controls.

Preprocessing

Standardized units and predefined primary endpoints.
Flow cytometry: compensation, locked gating strategy, arcsinh/log transformation.
Cytokines: values below detection limit imputed conservatively; log-transformed if skewed.
Imaging: predefined quantitative metrics (e.g., % procoagulant platelets, thrombin AUC).
TSPO binding: non-linear regression to determine Bmax and Kd.
Batch effects (if present) will be corrected statistically.

Statistical Analyses

Group comparisons: t-test/ANOVA (or Mann–Whitney/Kruskal–Wallis if non-normal).
Categorical variables: χ² or Fisher’s exact test.
Correlations: Spearman (primary).
Multivariable regression adjusting for age, sex, platelet count, and disease stage.
Multiple comparisons controlled using Benjamini–Hochberg FDR.
Effect sizes and 95% confidence intervals reported.

Hypothesis-Specific Analyses

H1: Platelets contain HCV RNA/proteins.
→ Compare platelet HCV positivity rates across stages; logistic regression for association with severity.

H2: HCV induces a hyperactivated/procoagulant platelet phenotype.
→ Compare activation markers, cytokines, EVs, and imaging endpoints across groups; correlate with viral measures.

H3: Specific receptors mediate platelet–HCV interaction.
→ Compare receptor expression across groups; test functional inhibition effects on viral binding and activation.

H4: TSPO expression is altered and linked to inflammation.
→ Compare TSPO density/expression across stages; correlate with cytokine levels and platelet activation markers.

Ethics and Data Management

This research proposal includes essential information regarding the involvement of human subjects in the study, demonstrating compliance with ethical standards and regulatory requirements. The study has received approval from the Institutional Review Board (IRB) of Cheikh Zaid Hospital in Rabat, Morocco, underscoring the commitment to safeguarding human participants' rights, safety, and well-being. The proposal outlines key elements pertaining to the human study, including protocols for obtaining informed consent, criteria for participant selection, and measures to ensure confidentiality and respect for participants' rights. The study also aligns with the principles of the Declaration of Helsinki, emphasizing its ethical foundation.

To address regulatory and ethical considerations, the proposal acknowledges the need to adhere to policies established by the Ministry of Public Health regarding protecting human subjects in research. It encourages examiners to refer to these policies for a comprehensive understanding of the regulatory framework governing the proposed research. As part of the ethical and regulatory framework, researchers are encouraged to consult with their institution's research compliance office before finalizing and submitting the proposal. This proactive step ensures that the research design, protocols, and ethical safeguards align with institutional and national standards.

In summary, the proposal provides transparent and thorough information about the human study, demonstrating compliance with ethical guidelines and emphasizing the importance of protecting the rights and well-being of research participants. The project will continue to adhere to all relevant ethical guidelines and regulatory requirements throughout its duration.

5. Budget

1. Personnel – $45,000

Postdoctoral Fellow (partial support) – $30,000

50% effort for 12 months
Responsible for experimental execution, data integration, supervision of students, and manuscript preparation.

Research Assistant / Technician (part-time) – $10,000

Sample processing, platelet isolation, assay preparation, data entry.

Graduate Student Support – $5,000

Research stipend supplementation and experimental support.

2. Laboratory Consumables – $25,000

Flow Cytometry Reagents & Antibodies – $10,000

Fluorochrome-conjugated antibodies (activation markers, receptors, EV markers)
Compensation beads, buffers, staining reagents

Cytokine Multiplex Kits / ELISA Kits – $6,000

Multiplex inflammatory panels
PF4, serotonin, and selected cytokine ELISAs

Molecular Biology Reagents – $4,000

RT-PCR reagents
RNA extraction kits
Primers and consumables

Cell Culture & Binding Assays Supplies – $3,000

Recombinant proteins
Inhibitory antibodies/peptides

General Laboratory Consumables – $2,000

Tubes, tips, buffers, isolation reagents

3. Imaging & Specialized Assays – $10,000

Confocal Imaging & Microscopy Core Fees – $5,000

Instrument access, imaging time, analysis software

Radioligand Binding / TSPO Assay Supplies – $5,000

Radioligand reagents
Membrane preparation materials

4. Biostatistics & Data Analysis – $5,000

Biostatistical consultation
Advanced modeling and validation
Data management support

5. Publication & Dissemination – $3,000

Open-access publication fees
Conference presentation support

6. Contingency / Pilot-Driven Adjustments – $2,000

Allocation reserved for adjustments based on early data
Additional reagents or validation assays if needed

Total Requested: $90,000 USD

Budget Flexibility Statement

These projected costs are estimates based on current experimental design and available pilot data. Allocation between personnel and consumables may be adjusted depending on interim findings (e.g., prioritizing receptor validation or TSPO mechanistic assays). The modular structure ensures efficient reallocation while maintaining scientific objectives.

6. References

1. Mohd Hanafiah K, Groeger J, Flaxman AD, Wiersma ST. Global epidemiology of hepatitis C virus infection: new estimates of age-specific antibody to HCV seroprevalence. Hepatology. 2013; 57(4):1333-42.

2. Cacoub P, Gragnani L, Comarmond C, Zignego AL. Extrahepatic manifestations of chronic hepatitis C virus infection. Dig Liver Dis. 2014; 46 Suppl 5:S165-73.

3. Boddi M, Abbate R, Chellini B, Giusti B, Giannini C, Pratesi G, et al. Hepatitis C virus RNA localization in human carotid plaques. J Clin Virol. 2010; 47(1):72-5.

4. Domont F, Cacoub P. Chronic hepatitis C virus infection, a new cardiovascular risk factor? Liver Int. 2016; 36(5):621-7.

5. Adinolfi LE, Zampino R, Restivo L, Lonardo A, Guerrera B, Marrone A, et al. Chronic hepatitis C virus infection and atherosclerosis: clinical impact and mechanisms. World J Gastroenterol. 2014; 20(13):3410-7.

6. Papadopoulos V, Baraldi M, Guilarte TR, Knudsen TB, Lacapere JJ, Lindemann P, et al. Translocator protein (18kDa): new nomenclature for the peripheral-type benzodiazepine receptor based on its structure and molecular function. Trends Pharmacol Sci. 2006; 27(8):402-9.

7. Sreeramkumar V, Adrover JM, Ballesteros I, Cuartero MI, Rossaint J, Bilbao I, et al. Neutrophils scan for activated platelets to initiate inflammation. Science. 2014; 346(6214):1234-8.

8. Fallahi P, Ferri C, Ferrari SM, Corrado A, Sansonno D, Antonelli A. Cytokines and HCV-related disorders. Clin Dev Immunol. 2012;2012:468107.

9. Morrell CN, Aggrey AA, Chapman LM, Modjeski KL. Emerging roles for platelets as immune and inflammatory cells. Blood. 2014; 123(18):2759-67.

10. Lindemann S, Tolley ND, Dixon DA, McIntyre TM, Prescott SM, Zimmerman GA, et al. Activated platelets mediate inflammatory signaling by regulated interleukin 1beta synthesis. J Cell Biol. 2001; 154(3):485-90.

11. Weizman A, Dagan E, Kaplan B, Karp L, Malmud S, Gil-Ad I, et al. Platelet peripheral-type benzodiazepine in pregnancy and lactation. Neuropsychopharmacology. 1999; 21(4):513-8.

12. Karas SP, Rosse WF, Kurlander RJ. Characterization of the IgG-Fc receptor on human platelets. Blood. 1982; 60(6):1277-82.

13. Idzko M, Pitchford S, Page C. Role of platelets in allergic airway inflammation. J Allergy Clin Immunol. 2015; 135(6):1416-23.

14. Imhof BA, Jemelin S, Ballet R, Vesin C, Schapira M, Karaca M, et al. CCN1/CYR61-mediated meticulous patrolling by Ly6Clow monocytes fuels vascular inflammation. Proc Natl Acad Sci U S A. 2016; 113(33):E4847-56.

15. Zaid Y, Puhm F, Allaeys I, Naya A, Oudghiri M, Khalki L, et al. Platelets Can Associate with SARS-Cov-2 RNA and Are Hyperactivated in COVID-19. Circ Res. 2020.

16. Ho-Tin-Noe B, Demers M, Wagner DD. How platelets safeguard vascular integrity. J Thromb Haemost. 2011; 9 Suppl 1:56-65.

17. Boulaftali Y, Hess PR, Kahn ML, Bergmeier W. Platelet immunoreceptor tyrosine-based activation motif (ITAM) signaling and vascular integrity. Circ Res. 2014; 114(7):1174-84.

18. Agbani EO, Van den Bosch MTJ, Brown E, Williams CM, Mattheij NJA, Cosemans JMEM, et al. Coordinated Membrane Ballooning and Procoagulant Spreading in Human Platelets. Circulation. 2015; 132(15):1414-24.

19. Agbani EO, Hers I, Poole AW. Temporal contribution of the platelet body and balloon to thrombin generation. Haematologica. 2017; 102(10):1-3.

20. Agbani EO, Williams CM, Li Y, van den Bosch MTJ, Moore SF, Mauroux A, et al. Aquaporin-1 Regulates Platelet Procoagulant Membrane Dynamics and In Vivo Thrombosis JCI Insight. 2018; 3(10):e99062.

21. Agbani EO, Williams CM, Hers I, Poole AW. Membrane Ballooning in Aggregated Platelets is Synchronised and Mediates a Surge in Microvesiculation. Scientific Reports. 2017; 7(2770):1-12.

22. Agbani EO, Schneider P, McDonald B, Skeith L, Poon M-C, Lee A. Activated Platelets Harbour SARS-CoV-2 During Severe COVID-19. Thromb Haemost. 2022; 122(2):308-9.

23. Marcoux G, Magron A, Sut C, Laroche A, Laradi S, Hamzeh-Cognasse H, et al. Platelet-derived extracellular vesicles convey mitochondrial DAMPs in platelet concentrates and their levels are associated with adverse reactions. Transfusion (Paris). 2019; 59(7):2403-14.

24. Boudreau LH, Duchez AC, Cloutier N, Soulet D, Martin N, Bollinger J, et al. Platelets release mitochondria serving as substrate for bactericidal group IIA-secreted phospholipase A2 to promote inflammation. Blood. 2014; 124(14):2173-83.

25. Monga S, Weizman A, Gavish M. The Efficacy of the Novel TSPO Ligands 2-Cl-MGV-1 and 2,4-Di-Cl-MGV-1 Compared to the Classical TSPO Ligand PK 11195 to Counteract the Release of Chemokines from LPS-Stimulated BV-2 Microglial Cells. Biology (Basel). 2020; 9(9).

26. Monga S, Nagler R, Amara R, Weizman A, Gavish M. Inhibitory Effects of the Two Novel TSPO Ligands 2-Cl-MGV-1 and MGV-1 on LPS-induced Microglial Activation. Cells. 2019; 8(5).

27. Azrad M, Zeineh N, Weizman A, Veenman L, Gavish M. The TSPO Ligands 2-Cl-MGV-1, MGV-1, and PK11195 Differentially Suppress the Inflammatory Response of BV-2 Microglial Cell to LPS. Int J Mol Sci. 2019; 20(3).

28. Lin YM, Sun HY, Chiu WT, Su HC, Chien YC, Chang HA, et al. Etifoxine, a TSPO Ligand, Worsens Hepatitis C-Related Insulin Resistance but Relieves Lipid Accumulation. Biomed Res Int. 2019;2019:3102414.

29. Liu M, Dou Y, Sun R, Zhang Y, Liu Y. Molecular mechanisms for alcoholic hepatitis based on analysis of gene expression profile. Hepat Mon. 2015; 15(5):e27336.

Platelets as Hidden Drivers of Hepatitis C Pathogenesis

Platelets as Hidden Drivers of Hepatitis C Pathogenesis

Funding Progress

Related Funding

Request for Proposals - Vascular Biology

Nonprofit Recipient

Topics

DOI