Diagnosis of Primary Ciliary Dyskinesia (PCD)

In parallel to our research in motile cilia we run a service at CEDOC- NMS to diagnose PCD. This disease requires a multidisciplinary approach to be correctly diagnosed as it can be mistaken with secondary causes such as respiratory infections. In coordination with the major Hospitals in the country we perform the analysis of the ciliated nasal epithelia of patients suspected to have PCD. Since 2014 that we offer the evaluation by high-speed video microscopy to quantify cilia beat frequency and beating pattern. Currently, we offer the electron microscopy study of the cilia ultrastructure in collaboration with the Southampton and London Hospitals (UK) and more recently we study the presence or absence of the proteins that confer motility to cilia by performing immunohistochemistry by immunofluorescence analysis. In the end we promote a multidisciplinary meeting where we join all the information from the different diagnostic techniques with the clinical history collected by clinicians and geneticists. The diagnostic of PCD is interactive and may involve collaboration with other Centres in Europe. Dr Susana Lopes is a supporting member of the PCD group of the ERN-LUNG European network where difficult cases are discussed among many clinicians and scientists.

Objective:
To diagnose patients with PCD and do research on PCD novel genes and translational research on potential therapies. In parallel with our investigation into motile cilia, we opened a PCD diagnostic service at CEDOC-NMS. 

Publications:

-CiliarMove: new software for evaluating ciliary beat frequency helps find novel mutations by a Portuguese multidisciplinary team on primary ciliary dyskinesia.
Sampaio P, da Silva MF, Vale I, Roxo-Rosa M, Pinto A, Constant C, Pereira L, Quintão CM, Lopes SS.
ERJ Open Res. 2021 Feb 8;7(1):00792-2020. doi: 10.1183/23120541.00792-2020. eCollection 2021 Jan.
PMID: 34104642 

-Zebrafish Motile Cilia as a Model for Primary Ciliary Dyskinesia.
Pinto AL, Rasteiro M, Bota C, Pestana S, Sampaio P, Hogg C, Burgoyne T, Lopes SS.
Int J Mol Sci. 2021 Aug 4;22(16):8361. doi: 10.3390/ijms22168361.
PMID: 34445067

-Primary ciliary dyskinesia due to CCNO mutations-A genotype-phenotype correlation contribution.
Henriques AR, Constant C, Descalço A, Pinto A, Moura Nunes J, Sampaio P, Lopes SS, Pereira L, Bandeira T.
Pediatr Pulmonol. 2021 Aug;56(8):2776-2779. doi: 10.1002/ppul.25440. Epub 2021 Jun 8.
PMID: 34102041 

1) 01-09-2013 to present time
Cilia and left-right establishment
Motile cilia are responsible for many processes in vertebrate development, one of the most fascinating being the determination of the left and right sides of the body. Since 2010 that we have been investigating how the left-right axis is first laid down. Special motile cilia are present in monociliated cells lining the left-right organizer, a transient organ present in most vertebrates including humans. We had 3 FCT funded projects on this subject.

2)  01-09-2015 to present time
Recover motile cilia function
We have special interest in the disease Primary Ciliary Dyskinesia (PCD). Our lab investigates the causes of this disease using the zebrafish olfactory pit, an organ with multiciliated cells that recapitulates the human respiratory epithelia. Our studies involve using formulated mRNA to restore cilia motility and we will start a new project on this subject in 2022 funded by Maratona da Saúde.

3) 01-01- 2014 to 01-08-2021
How can zebrafish help to investigate Polycystic Kidney Disease?
Cilia have acquired major biomedical relevance due to the disclosure of many different diseases as ciliopathies i.e. diseases that have a ciliary origin, or somehow involve the cilium, such as Autosomal Dominant Polycystic Kidney Disease (ADPKD). In this project we have demonstrated that the left right organizer of the zebrafish can be used as a model for a kidney cyst. We had funding from FCT and from Portuguese Society of Nephrology.

4) 01-01-2014 to present time
Diagnostics of Primary Ciliary Dyskinesia (PCD)
In parallel to our research in cilia we run a service at CEDOC- NMS to diagnose PCD. This disease requires a multidisciplinary approach to be correctly diagnosed as it can be mistaken with secondary causes such as respiratory infections. In coordination with the major Hospitals in the country we perform the analysis of the ciliated nasal epithelia of patients suspected to have PCD. Since 2014 that we offer the evaluation by high-speed video microscopy to quantify cilia beat frequency and beating pattern. Currently we also offer the electron microscopy study of the cilia ultrastructure in collaboration with the Southampton and London PCD services, UK). More recently, we study the presence or absence of the proteins that confer motility to cilia by performing immunohistochemistry by immunofluorescence analysis. Along with the diagnostic service we do research on PCD like the development of new open source software for cilia frequency evaluation.

Zebrafish Model as a Screen to Prevent Cyst Inflation in Autosomal Dominant Polycystic Kidney Disease.
Oliveira I, Jacinto R, Pestana S, Nolasco F, Calado J, Lopes SS, Roxo-Rosa M.
Int J Mol Sci. 2021 Aug 20;22(16):9013. doi: 10.3390/ijms22169013.
PMID: 34445719 
 
Pkd2 Affects Cilia Length and Impacts LR Flow Dynamics and Dand5.
Jacinto R, Sampaio P, Roxo-Rosa M, Pestana S, Lopes SS.
Front Cell Dev Biol. 2021 Apr 1;9:624531. doi: 10.3389/fcell.2021.624531. eCollection 2021.
PMID: 33869175

Imbalanced mitochondrial function provokes heterotaxy via aberrant ciliogenesis.
Burkhalter MD, Sridhar A, Sampaio P, Jacinto R, Burczyk MS, Donow C, Angenendt M; Competence Network for Congenital Heart Defects Investigators, Hempel M, Walther P, Pennekamp P, Omran H, Lopes SS, Ware SM, Philipp M.
J Clin Invest. 2019 May 16;129(7):2841-2855. doi: 10.1172/JCI98890.
PMID: 31094706

Notch/Her12 signalling modulates, motile/immotile cilia ratio downstream of Foxj1a in zebrafish left-right organizer.
Tavares B, Jacinto R, Sampaio P, Pestana S, Pinto A, Vaz A, Roxo-Rosa M, Gardner R, Lopes T, Schilling B, Henry I, Saúde L, Lopes SS.
Elife. 2017 Sep 6;6:e25165. doi: 10.7554/eLife.25165.
PMID: 28875937

Dynamics of cilia length in left-right development.
Pintado P, Sampaio P, Tavares B, Montenegro-Johnson TD, Smith DJ, Lopes SS.
R Soc Open Sci. 2017 Mar 8;4(3):161102. doi: 10.1098/rsos.161102. eCollection 2017 Mar.
PMID: 28405397 

Left-right organizer flow dynamics: how much cilia activity reliably yields laterality?
Sampaio P, Ferreira RR, Guerrero A, Pintado P, Tavares B, Amaro J, Smith AA, Montenegro-Johnson T, Smith DJ, Lopes SS.
Dev Cell. 2014 Jun 23;29(6):716-28. doi: 10.1016/j.devcel.2014.04.030. Epub 2014 Jun 12.
PMID: 24930722

Maratona da Saúde Senior prize on Rare Diseases 2021

Professor Heymut Omran, University of Muenster, Germany 
Professor David Smith, University of Birmingham, UK
Ethris GmbH, Germany