Dr Harprit Singh

Job: Faculty Head of Research Students, Associate Professor/Reader

Faculty: Health and Life Sciences

School/department: School of Allied Health Sciences

Address: De Montfort University, The Gateway, Leicester, LE1 9BH.

T: +44 (0)116 257 7779

E: harprit.singh@dmu.ac.uk

W: https://www.dmu.ac.uk/research/centres-institutes/iahsr/index.aspx

 

Personal profile

An active non-clinical researcher with interests in the area of vascular biology and metabolic disease. Primary focus of research is around endothelial cell signalling involved in vascular stability. Lead the Nutrition and Metabolic Health research theme within the Institute of Allied Health Sciences Research. 

As Faculty Head of Research Students lead and support Postgraduate Research Students with their research degree completions.

Research group affiliations

Publications and outputs


  • C2- linked alkynyl poly-ethylene glycol(PEG) adenosine conjugates as water-soluble adenosine receptor agonists
    C2- linked alkynyl poly-ethylene glycol(PEG) adenosine conjugates as water-soluble adenosine receptor agonists Ferguson, Lindsay; Madieh, Nasrin Shokrzadeh; Brucoli, Federico; Vaideanu, Alexandra; Schatzlein, Andreas; Festa, Joseph; Singh, Harprit; Wells, Geoffrey; Bhakta, Sanjib A series of 12 novel polyethylene-glycol(PEG)-alkynyl C2-adenosine(ADN) conjugates were synthesized using a robust Sonogashira coupling protocol and characterized by NMR spectroscopy and mass spectrometry analysis. The ADN-PEG conjugates showed null to moderate toxicity in murine macrophages and 12c was active against Mycobacterium aurum growth (MIC = 62.5 mg/L). The conjugates were not active against Mycobacterium bovis BCG. Conjugates 10b and 11b exhibited high water solubility with solubility values of 1.22 and 1.18 mg/ml, respectively, in phosphate buffer solutions at pH 6.8. Further, 10b and 11b induced a significant increase in cAMP accumulation in RAW264.7 cells comparable with that induced by adenosine. Analogues 10c, 11c and 12c were docked to the A1, A2A, A2B and A3 adenosine receptors (ARs) using crystal-structures and homology models. ADN-PEG-conjugates bearing chains with up to five ethyleneoxy units could be well accommodated within the binding sites of A1, A2A and A3 ARs. Docking studies showed that compound 10b and 11b were the best A2A receptor binders of the series, whereas 12c was the best binder for A1 AR. In summary, introduction of hydrophilic PEG substituents at the C2 of adenine ring significantly improved water solubility and did not affect AR binding properties of the ADN-PEG conjugates. open access article Ferguson, L., Madieh, N.S., Vaideanu, A., Schatzlein, A., Festa, J., Singh, H., Wells, G., Bhakta, S., Brucoli, F. (2022) C2‐linked alkynyl poly‐ethylene glycol (PEG) adenosine conjugates as water‐soluble adenosine receptor agonists. Chemical Biology & Drug Design.
  • Elderberries as a potential supplement to improve vascular function in a SARS-CoV-2 environment
    Elderberries as a potential supplement to improve vascular function in a SARS-CoV-2 environment Festa, Joseph; Singh, Harprit; Hussain, Aamir; Da Boit, Mariasole Coronavirus disease 2019 (COVID-19) pandemic has been triggered by the severe acute respiratory syndrome coronavirus (SARS-CoV-2). Although recent studies demonstrate that SARS-CoV-2 possibly does not directly infect endothelial cells (EC), the endothelium may be affected as a secondary response due to the damage of neighboring cells, circulating pro-inflammatory cytokines, and/or other mechanisms. Long-term COVID-19 symptoms specifically nonrespiratory symptoms are due to the persistence of endothelial dysfunction (ED). Based on the literature, anthocyanins a major subgroup of flavonoid polyphenols found in berries, have been well researched for their vascular protective properties as well as the prevention of cardiovascular disease (CVD)-related deaths. Elderberries have been previously used as a natural remedy for treating influenza, cold, and consequently cardiovascular health due to a high content of cyanidin-3-glucoside (C3G) a major anthocyanin found in the human diet. The literature reported many studies demonstrating that EE has both antiviral and vascular protective properties that should be further investigated as a nutritional component used against the (in)direct effect of SARS-CoV-2 in vascular function. The Publisher's final version can be found by following the DOI link.. Open access article. Festa, J., Singh, H., Hussain, A. and Da Boit, M. (2022) Elderberries as a potential supplement to improve vascular function in a SARS-CoV-2 environment. Journal of Food Biochemistry, 00, e14091.
  • Synthesis and Identification of Biologically Active Mono-Labelled FITC-Insulin Conjugate
    Synthesis and Identification of Biologically Active Mono-Labelled FITC-Insulin Conjugate Sahota, Tarsem; Vu, Tam; Taylor, M Joan; Singh, Harprit; Bottrill, Andrew; Bilmoria, Jay Fluorescently labelling proteins such as insulin have wide ranging applications in a pharmaceutical research and drug delivery. Human insulin (Actrapid®) was labelled with fluorescein isothiocyanate (FITC) and the synthesised conjugate identified using reverse phase high performance liquid chromatography (RP-HPLC) on a C18 column and a gradient method with mobile phase A containing 0.1% trifluoroacetic acid (TFA) in Millipore water and mobile phase B containing 90% Acetonitrile, 10% Millipore water and 0.1% TFA. Syntheses were carried out at varying reaction times between 4 and 20 h. Mono-labelled FITC-insulin conjugate was successfully synthesised with labelling at the B1 position on the insulin chain using a molar ratio of 2:1 (FITC:insulin) at a reaction time of 18 h and confirmed by electrospray mass spectroscopy. Reactions were studied across a pH range of 7–9.8 and the quantities switch from mono-labelled to di-labelled FITC-insulin conjugates at a reaction time of 2 h (2:1 molar ratio) at pH > 8. The conjugates isolated from the studies had biological activities in comparison to native insulin of 99.5% monoB1, 78% monoA1, 51% diA1B1 and 0.06% triA1B1B29 in HUVEC cells by examining AKT phosphorylation levels. MonoB1 FITC-insulin conjugate was also compared to native insulin by examining cell surface GLUT4 in C2C12 skeletal muscle cells. No significant difference in the cellular response was observed for monoB1 produced in-house compared to native insulin. Therefore mono-labelled FITC-insulin at the B1 position showed similar biological activity as native insulin and can potentially be used for future biomedical applications. The file attached to this record is the author's final peer reviewed version. The Publisher's final version can be found by following the DOI link. Vu, T., Taylor, M.J., Singh, H., Bottrill, A., Bilmoria, J., Sahota, T. (2022) Synthesis and Identification of Biologically Active Mono-Labelled FITC-Insulin Conjugate. Journal of Fluorescence, 32, pp. 569-582
  • Potential Benefits of Berry Anthocyanins on Vascular Function
    Potential Benefits of Berry Anthocyanins on Vascular Function Festa, Joseph; Hussain, Aamir; Da Boit, Mariasole; Singh, Harprit The file attached to this record is the author's final peer reviewed version. The Publisher's final version can be found by following the DOI link. Festa, J., Da, M., Hussain, A., Singh, H. (2021) Potential Benefits of Berry Anthocyanins on Vascular Function. Molecular Nutrition and Food Research, 2100170. https://doi.org/10.1002/mnfr.202100170
  • BS20 The opposing effects of chronic interleukin-1β on TIE2:TIE1 ratio and angiopoietin1 induced PI3 Kinase/AKT signaling in endothelial cells
    BS20 The opposing effects of chronic interleukin-1β on TIE2:TIE1 ratio and angiopoietin1 induced PI3 Kinase/AKT signaling in endothelial cells Singh, Harprit; Bilimoria, Jay Introduction Angiopoietin-1 (Ang-1) is a growth factor that plays a crucial role in maintaining normal vascular function. The main role of Ang-1 is to maintain endothelial survival. Ang-1 exerts its protective effect by activating Tie-2 receptors and subsequently the Phosphatidylinositol 3-kinase (PI3Kinase)/ AKT pathway. Tie-1 regulates Ang-1 signalling with high levels of the receptor reducing Ang-1-induced Tie-2 activation. Proinflammatory cytokines including Tumor Necrotic factor (TNF-a) Interleukin 1b (IL-1b) have been implicated in a range of vascular pathologies including vascular inflammation and atherosclerosis. While TNF-a has shown to regulate Tie receptors, the chronic impact IL-1b has on Ang-1/Tie receptor signalling pathway and vascular function has not been investigated. Aim To examine the impact IL-1b has on Tie receptor expression and Angiopoietin-1 induced PI3Kinase/AKT activity in endothelial cells. Method Primary Human Umbilical Vein Endothelial Cells (HUVEC) were stimulated with 25ng/ml of IL-1b in the presence or absence of 100ng/ml of human recombinant Ang-1. The treatment times ranged from 0 to 48h. Cell lysates from the treated cells were then subjected to Western blotting to analyze Tie receptor levels and phospho-AKT (pAKT), a signaling molecule associated with Ang-1 cellular transduction. The levels of target proteins were compared between reactions by quantifying mean intensity of bands. In addition, cells were treated with Ang-1 in the absence or presence of IL-1b at various time points. The cell viability assay was performed on the treated cells by following the manufactures protocol. The mean percentage of live to dead cells was calculated from three random fields for each treatment. Data for the Tie receptor and AKT analysis is presented as means and SEM of three independent experiments. Statistical significance represented with p<0.05 using Student’s t-test. Data for the cell viability assay is presented as means and SEM of two independent experiments. Results A significant reduction in the levels of Tie-1 receptor was observed at 3h (58.6±8.6%) in HUVECs treated with IL1b. The cytokine was able to maintain significant low levels of Tie1 up until 48h of treatment, whereas the changes in levels of Tie-2 were insignificant. Interestingly, IL-1b significantly reduced Ang1-induced pAKT activity from 3h onwards with maximum reduction of 73.4±12.8% observed at 48h. The cell viability assays showed reduction in the percentage live to dead cells between Ang-1 and Ang1 + IL-1b for chronic time points tested. Conclusion Long term exposure of Interleukin-1b is capable of altering Tie-1 levels and increasing the Tie-2: Tie-1 ratio in endothelial cells. In contracts, IL-1b reduces Ang-1-PI3Kinase/ AKT signalling and endothelium cell viability. This opposing phenomenon observed where IL-1b impairs Ang-1 protective ability suggests a different mechanism of regulation, independent of the Tie-1 receptor. Singh, H., Bilimoria, J. (2021) BS20 The opposing effects of chronic interleukin-1β on TIE2:TIE1 ratio and angiopoietin1 induced PI3 Kinase/AKT signaling in endothelial cells. British Cardiovascular Society Annual Conference, 7-10th June 2021, Virtual.
  • The use of near-peer tutors to improve level of learning & confidence in areas of human physiology
    The use of near-peer tutors to improve level of learning & confidence in areas of human physiology Iqbal, Fatima; Gaffar, Fatema; Singh, Harprit This study investigates the impact near-peer tutors (NPTs) have on the learning experience of students enrolled on the biomedical and medical science programmes at De Montfort University, UK. Using a questionnaire, tutees were able to comment on their confidence level on certain academic taught topics in human physiology before and after attending near-peer tutored sessions. Statistical analysis showed a significant increase in the level of confidence observed by the tutees after attending the sessions (before vs after, p < 0.05). Tutees enjoyed the level of interaction and active learning with huge praise towards the NPTs. Qualitative data were also captured on the experience and skills gained by NTPs. The near-peer tutors highlighted the importance of these sessions in enhancing their knowledge and gaining valuable employability skills for the workplace. In conclusion, supporting lecturers with additional student-led sessions improved confidence levels and understanding of topics in human physiology among tutees. Having similar NPTs sessions across programmes can prove to be beneficial to student engagement & learning in higher education The file attached to this record is the author's final peer reviewed version. The Publisher's final version can be found by following the DOI link. Iqbal, F., Gaffar, F. and Singh, H. (2020) The use of near-peer tutors to improve level of learning & confidence in areas of human physiology. Journal of Biological Education,
  • The Angiopoietin ligands and Tie receptors: potential diagnostic biomarkers of vascular disease
    The Angiopoietin ligands and Tie receptors: potential diagnostic biomarkers of vascular disease Bilimoria, Jay; Singh, Harprit The Angiopoietin-1 (Angpt1)/Tie2 signaling pathway is important in regulating vascular function. Angpt1-induced Tie2 activation promotes vascular endothelial cell survival and reduces vascular leakage. Angiopoietin-2 (Angpt2), a weak agonist/antagonist of Tie2, opposes and regulates Angpt1 action. The Tie family of receptor tyrosine kinases, Tie2 and Tie1, exist as either homo-or heterodimers. The molecular complex between the receptors is also crucial in controlling Angpt1 signaling; hence, the molecular balance between Angpt1:Angpt2 and Tie2:Tie1 is important in determining endothelial integrity and vascular stability. This review presents evidence of the change observed in the Angiopoietin/Tie molecules in various pathophysiological conditions and discusses the potential clinical applications of these molecules in vascular complications. The file attached to this record is the author's final peer reviewed version. The Publisher's final version can be found by following the DOI link. Bilimoria, J. and Singh, H. (2019) The Angiopoietin ligands and Tie receptors: potential diagnostic biomarkers of vascular disease, Journal of Receptors and Signal Transduction,
  • Effects of novel chalcone derivatives on human endothelial cell proliferation and migration
    Effects of novel chalcone derivatives on human endothelial cell proliferation and migration Hussain, A.; Fretwell, L.; Ruparelia, K. C.; Beresford, Kenneth J. M.; Singh, Harprit Introduction: Angiogenesis is the formation of new blood vessels from the pre-existing vasculature and is essential for some physiological processes including wound healing and menstrual cycles. Unregulated angiogenesis can lead to vascular related diseases such as age related macular degeneration, cancer and rheumatoid arthritis. Chalcones (1,3-diphenylpropenones) are naturally occurring phenolic compounds found in a variety of plants and fruits. Their simple molecular structures have demonstrated an array of pharmacological activity including antioxidant and anti-vascular. Due to Chalcones having an easily modifiable scaffold, they are widely used as parent compounds in drug discovery studies. Endothelial cell proliferation and migration are essential for angiogenic growth, so compounds that possess anti-endothelial activity could be developed further as potential inhibitors of angiogenesis. With this in mind, we report the anti-endothelial activity of two novel analogues (AH1 and AH9) derived from the parent compound 2-chloro-2’5’-dihydroxychalcone. Method: Chalcones were synthesised via the Claisen-Schmidt condensation and verified via nuclear magnetic resonance (NMR) and mass spectrometry (MS). Primary Human umbilical vein endothelial cells (HUVECs) were cultured according to manufactures protocol. Cells were seeding into 96-well plates (3500 cells/well) and, 24 hours later, treated either with AH1, AH9 or Sorafenib, a known anti-angiogenic drug for 72hr at a concentration of 10 µM. Effects of the compounds on cell proliferation was assessed by quantifying the colorimetric conversion of 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT) to purple formazan product (measured at 560 nm). Data are expressed as mean % inhibition of cell proliferation ± SEM (n=4). Anti-migratory activity was assessed via the wound healing assay and the effects of the compounds at 3 µM on the width of the scratch across an 8 hour period were compared to untreated control. Analysis was performed by one-way ANOVA with Tukey-Kramer’s multiple comparisons test. Data are expressed as mean % of maximum migration ± SEM (n=3). Results: AH1 and AH9 displayed significant anti-proliferative activity with inhibitory values of 94.94 ± 1.64% and 79.62 ± 4.45% compared to Sorafenib, 69.95 ± 4.12%, respectively. Effects on HUVEC migration showed that AH9 limited migration to 16.19 ± 14.44% (P<0.05 vs. control). AH1 (47.23 ± 6.8%) showed similar anti-migratory activity to Sorafenib (52.68 ± 3.32%). Conclusion: This preliminary data suggests that AH9 has potential antiangiogenic properties and could be developed further as a potential antiangiogenic or vascular remodelling agent. Further studies will elucidate the molecular mechanism of AH9 action in cells stimulated with known angiogenic agent, VEGF. Hussain, A., Fretwell, L., Ruparelia, K., Beresford, K., Singh, H.(2018) Effects of novel chalcone derivatives on human endothelial cell proliferation and migration. Cardiovascular Research, 114 (Supplement 1), pp.S50-S51.
  • Biomedical science students as teachers to levels KS3 and KS4
    Biomedical science students as teachers to levels KS3 and KS4 Pena-Fernandez, A.; Pena, M. A.; Lobo-Bedmar, M. C.; Singh, Harprit; Weldon, C. Students-as-teachers is a useful pedagogy for future professionals that require teaching abilities in their profession such as nurses and medical doctors. However, this has been little explored in other human health sciences programmes such as biomedical (BMS) and medical science (BMedS) degrees. A novel pedagogy has been introduced at De Montfort University (UK) to develop teaching abilities in second and final year students enrolled in BMS and BMedS programmes. A total of 15 students were deployed to Bermuda in November 2016 for 2 weeks to teach different topics of human biology and microbiology to secondary school children level KS3 (aged 11-14) and KS4 (aged 14-18) with academic-supervision. Briefly, this pedagogy consisted of students distributed into groups of 5 to encourage peer-interaction, with one group (the teaching group) responsible for leading different teaching/learning sessions in microbiology (e.g. virus and bacteria, disease, hygiene, prokaryotic and eukaryotic cells, etc.). The other groups peer-assessed the performance of the teaching group in conjunction with the academics. This approach facilitated the provision of comprehensive feedback so students could improve their performance throughout their deployment. 73% of students enjoyed teaching and 100% highlighted that were highly satisfied with the experience. 29% would like to do a teaching career (40% of students reported uncertainty about their future career); however, 80% of students highlighted that this experience will help to decide their career. Only 13% students reported that they struggled in preparing their topics, particularly to KS3. Finally, students indicated that the best part of the experience was inspiring young people and when schoolchildren provided feedback and were interactive. Confidence was perceived as a skill to improve. Students reported high levels of satisfaction and recommended the inclusion of similar pedagogies in their degrees. We consider that involving students in teaching provide some teaching abilities and critical transversal competences. Peña-Fernández A., Peña MA., Lobo-Bedmar MC., Singh H., Weldon C. (2017) Biomedical science students as teachers to levels KS3 and KS4. The Institute of Biomedical Science (IBMS) Congress, Birmingham, UK, September 2017.
  • The effect of Interleukin 1 Beta on vascular Angiopoietin 1 signalling
    The effect of Interleukin 1 Beta on vascular Angiopoietin 1 signalling Bilimoria, Jay; Singh, Harprit Bilimoria, J. and Singh, H. (2017) The effect of Interleukin 1 beta on vascular Angiopoietin 1 signalling. The Annual British Society For Cardiovascular Research/British Cardiovascular Society Conference, Manchester, June 2017

Click here to view a full listing of Dr Harprit Singh's publications and outputs 

Research interests/expertise

Involved in investigating the mechanism associated with metabolic syndrome induced endothelial dysfunction and cardiovascular disease in patients. Research expertise include:

  • Cardiovascular Disease
  • Diabetes
  • Endothelial biology
  • Cell signalling
  • Growth hormones and their Receptors.
  • Molecular markers of vascular disease
  • Translational research

Areas of teaching

  • Endocrinology
  • Cardiovascular 
  • Human Anatomy and Physiology
  • Angiogenesis

Qualifications

  • PhD in Biochemistry
  • PGCert in Teaching and Learning in Higher Education
  • BSc (Hons) Medical Biochemistry

Courses taught

  • Medical Science (BMed Sci)
  • Biomedical Science (BSc)
  • Adv Biomedical Science (MSc)

Honours and awards

  • Fellow of the Higher Education Academy (FHEA)

Membership of professional associations and societies

  • The British Society for Cardiovascular Research
  • European Society of Cardiology Working Group Atherosclerosis & Vascular Biology

Current research students

  • Joseph Festa, PhD
  • Koddus Ali, PhD
  • Nasrin Shokrzadeh, PhD 
  • Tam Nqoc Minh Vu, PhD

Externally funded research grants information

 

Potential PhD projects for students

  • Diabetes and Endothelial signalling
  • Impact of Polyunsaturated fat on Endothelial signalling
  • Predicting vascular disease

Successfully Supervised PhD Students

 Jay Bilimoria, 2016-2020

Aamir Hussain, 2015-2019

 Rasmieh Alziedan, 2012-2016

 

Journal Peer Reviewer

  • Frontiers in Physiology: Vascular Physiology
  • Journal of Biomedical Research
  • Cancer Letters
  • Human Cell

Grant Peer Reviewer

Medical Research Council