Cell Communication in Disease Pathology

Blood diseases affecting red blood cells

This ongoing research has two objectives:

1. Extracellular vesicles in malaria pathology and diagnosis

Malaria-infected red blood cells release tiny particles called extracellular vesicles that worsen disease, and our previous work shows these vesicles can be detected in the blood very early in infection. This project aims to reduce malaria severity by blocking vesicle release from red cells and to improve early diagnosis by identifying vesicle-based biomarkers in patient samples.

Together with our partners at the University of Ghana (Dept of Medical Laboratoy Sciences), we have described a pathogenic effect of RBC EVs from malaria-infected mice. EVs therefore play a significant role in the health of RBCs during malaria infection.

The aim of the project is to inhibit EV biogenesis from red cells in infection to ameliorate disease pathology. From a large cross-sectional field study we have also previously found plasma EVs to be early indicators of infection, before detectable parasitaemia levels (Antwi-Baffour, S. et al., J Extracell Vesicles 2019 Dec 18;9(1):1697124) and are now proceeding with further field studies to detect particular miRNA markers in plasma EVs from malaria patients.

2. Extracellular vesicles and cfDNA in sickle cell disorder (SCD)

This project will improve how extracellular vesicles and DNA fragments in the blood are measured in people with SCD and examine how these change during painful vaso-occlusive crises. By linking these blood markers to inflammation, immune cell activation, and crisis severity, the study aims to identify better ways to understand, monitor, and potentially predict severe disease episodes.

In this project in collaboration with the Whittington Health NHS Trust, we are optimising laboratory protocols for isolating EVs in SCD and distinguishing EVs from erythroid, endothelial and leukocyte populations. We are further comparing analytical platforms to define the most informative approaches for EV characterization in SCD.

We aim to establish an in vitro platform to evaluate the functional impact of SCD-derived EVs on immune cells focusing on macrophage activation, and inflammatory signalling. The project will determine whether EV subpopulations and cargo change significantly during Vaso-occlusive crises (VOCs) and ascertain whether these characteristics correlate with VOC severity. This study will also investigate whether EVs modulate macrophage response, contributing to immune dysregulation in SCD.

We will also investigate cell-free DNA (cfDNA). Having defined the minimum blood volume for robust cfDNA quantification and downstream analysis, and having differentiated nuclear from mitochondrial cfDNA, this project will investigate their relative contribution to disease severity and the influence of sampling timepoint on cfDNA levels during VOCs. As with EVs, but now relating to cfDNA, their levels and methylation patterns will also be correlated with VOC severity.

1. Needham SR, Davis BM, Uysal-Onganer P, Rolfe DJ, Hristova M, Kraev I, Inal JM, Lange S. (2025) Profiling Protein Citrullination in Extracellular Vesicles by Single-Molecule Detection Using Direct Stochastic Optical Reconstruction Microscopy. J Biophotonics. 2025 Nov 27:e202500483. 
2. Mannaperuma, D. Stratton, D., Lange, S. and Inal, J.M. (2025) Extracellular Vesicles from the Myocyte Secretome Contribute In Vitro to Creating an Unfavourable Environment for Migrating Lung Carcinoma Cells. Biology 14(11), 1578.
3. Lange, S., Bernstein, D.E., Dimov, N., Puttaswamy, S., Johnston, I., Kraev, I., Needham, S.R., Vasdev, N., Inal, J.M. (2025) Urinary extracellular vesicle signatures as biomarkers in prostate cancer patients. International journal of molecular sciences 26 (14), 6895
4. Smarslik, M., Inal, J.M. (2025) Synergistic effect of 5-fluorouracil and the small molecule Wnt/β-catenin inhibitor iCRT3 on Caco-2 colorectal cancer cells in vitro. Heliyon 11 (10)
5. Hearfield, N., Brotherton, D., Gao, Z., Inal, J., Stotz, H.U. (2025) Establishment of an experimental system to analyse extracellular vesicles during apoplastic fungal pathogenesis. Journal of Extracellular Biology 4 (2), e70029
6. Ansa-Addo, E.A., Pathak, P., McCrossan, M.V, Rossi, I., V., Abdullahi, M., Stratton, D., Lane, S., Ramirez, M.I., Inal, J.M. (2024) Monocyte-derived Extracellular Vesicles, stimulated by Trypanosoma cruzi, enhance cellular invasion in vitro, via activated TGF-β1. J Extracell Vesicles. 13(11):e70014.
7. Lange, S., Inal, J.M., Kraev, I., Alwyn Dart, D., and Uysal-Onganer, P. (2024) Low Magnetic Field Exposure alters Prostate Cancer Cell Properties. Biology 13(9), 734.
8. Welsh, J. A., Goberdhan, D. C. I., O’Driscoll, L., Buzas, E. I., Blenkiron, C., Bussolati, B., Cai, H., Di Vizio, D., Driedonks, T. A. P., Erdbrügger, U., Falcon-Perez, J. M., Fu, Q.-L., Hill, A. F., Lenassi, M., Lim, S. K., Mahoney, M. G., Mohanty, S., Möller, A., Nieuwland, R., … Witwer, K. W. (2024). Minimal information for studies of extracellular vesicles (MISEV2023): from basic to advanced approaches. Journal of Extracellular Vesicles, 13, e12404.
9. Goh, S. and Inal, J.M. (2024) Membrane vesicles of Clostridioides difficile and other Clostridial species. In series: Advances in Microbiology, Infectious Diseases and Public Health. Book title: Updates on Clostridioides difficile in Europe, Vol 4136, pp315-327, Springer Nature.
10. Lange, S. and Inal, J.M. (2023) Animal Models of Human Disease. Int. J. Mol. Sci. 24(21), 15821
11. Jorfi, S., Ansa-Addo, E.A., Mariniello, K., Warde, P., Bin Senian, A.A., Stratton, D., Bax, B.E., Levene, M., Lange, S., and INAL, J.M. (2023). A Coxsackievirus B1-mediated nonlytic Extracellular Vesicle-to-cell mechanism of virus transmission and its possible control through modulation of EV release. J Gen Virol. 104(9).
12. Rossi, I.V., Nunes, M.A.F., Sabatke, B., Ribas, H.T., Winnischofer S.M.B., Ramos, A.S.P., INAL, J.M., and Ramirez, M.I. (2022) An induced population of Trypanosoma cruzi epimastigotes more resistant to complement lysis promotes a phenotype with greater differentiation, invasiveness and release of extracellular vesicles. Front. Cell. Infect. Microbiol. 12:1046681.
13. Inal, J. Paizuldaeva A, Terziu E. (2022) Therapeutic use of calpeptin in COVID-19 infection. Clin Sci (Lond). Oct 28;136(20):1439-1447
14. Stratton, D. Malibha-Pinchbeck, M. and Inal, J. (2022) Extremely low-frequency magnetic fields significantly enhance the cytotoxicity of methotrexate and can reduce migration of cancer cell lines via transiently induced plasma membrane damage. Biochem. Biophys. Res. Commun. 626, 192-199
15. Inal, J.M., Hristova, M., and Lange, S. (2022) A Pilot Study on Peptidylarginine Deiminases and Protein Deimination in Animal Cancers across Vertebrate Species. Int. J. Mol. Sci. 23(15):8697.
16. De Sousa K.P., Rossi, I., Abdullahi, M., Ramirez, M.I., Stratton, D., and Inal, J.M. (2022) Isolation and characterization of extracellular vesicles and future directions in diagnosis and therapy. WIREs Nanomedicine & Nanobiotechnology Jul 27:e1835.
17. Laich A, Patel H., Zarantonello A., Sim, R.B., Inal, J.M. (2022) C2 by-pass: Cross-talk between the complement classical and alternative pathways. Immunobiology. 227(3):152225.
18. Stotz, H.U., Brotherton, D. and Inal, J.M. (2022) Communication is key: Extracellular vesicles as mediators of infection and defence during host-microbe interactions in animals and plants. FEMS Microbiol. Rev.46(1)fuab044.
19. Bernstein DE, Piedad J, Hemsworth L, West A, Johnston ID, Dimov N, Inal, J.M., and Vasdev N. (2021) Prostate cancer and microfluids. Urol Oncol. S1078-1439(21)00120-4.
20. Inal, J. (2020) Biological Factors Linking ApoE ε4 Variant and Severe COVID-19. Curr Atheroscler Rep.(11):70.
21. Inal, J. (2020) Complement-mediated Extracellular Vesicle release as a measure of endothelial dysfunction and prognostic marker for COVID-19 in peripheral blood - Letter to the Editor. Clin Hemorheol Microcirc. 2020;75(4):383-386.
22. Inal, J. (2020) COVID-19 comorbidities, associated procoagulant extracellular vesicles and venous thromboembolisms: a possible link with ethnicity? Br J Haematol. 10.1111
23. Inal J.M. (2020) Decoy ACE2-expressing extracellular vesicles that competitively bind SARS-CoV-2 as a possible COVID-19 therapy. Clin Sci (Lond). 134(12):1301-1304.
24. Uysal-Onganer, P., MacLatchy, A., Mahmoud, R., Kraev, I., Thompson, P.R., Inal, J.M., Lange, S. (2020) Peptidylarginine deiminase isozyme-specific PAD2, PAD3 and PAD4 inhibitors differentially modulate extracellular vesicle signatures and cell invasion in two glioblastoma multiforme cell lines. International Journal of Molecular Sciences 21(4). pii: E1495
25. Antwi-Baffour, S., Malibha-Pinchbeck, M., Stratton, D., Lange, S., Inal, J.M. (2019) Plasma mEV levels in Ghanain malaria patients with low parasitaemia are higher than those of healthy controls, raising the potential for parasite markers in mEVs as diagnostic targets. J. Extracell. Vesicles 9(1):1697124
26. Kosgodage U.S., Matewele P., Awamaria B., Kraev I., Warde P., Mastroianni G., Nunn A.V., Guy G.W., Bell J.D., Inal, J.M., Lange S. (2019) Cannabidiol Is a Novel Modulator of Bacterial Membrane Vesicles. Front Cell Infect Microbiol. 10;9:324.
27. Kosgodage U., Matewele P, Mastroianni G, Kraev I, Brotherton D, Awamaria B, Nicholas AP, Lange S, and Inal J.M. (2019) Peptidylarginine Deiminase Inhibitors Reduce Bacterial Membrane Vesicle Release and Sensitize Bacteria to Antibiotic Treatment. Front Cell Infect Microbiol. 9:227.
28. Sisa, C. Kholia, S., Naylor, J., Herrera Sanchez, M., Bruno, S., Deregibus, M., Camussi, G., Inal, J.M., Lange, S., Hristova, M. (2019) Mesenchymal Stem-Cell derived Extracellular Vesicles reduce Hypoxia-Ischemia Induced Perinatal Brain Injury. Frontiers Physiol. 10:282.