Spatial Analysis of Host-parasite Interactions in Cutaneous Leishmaniasis in Ethiopia (SpatialCL)

• ClinicalTrials.gov Identifier: NCT05332093
• Recruitment Status: Recruiting
• First Posted: April 18, 2022
• Last Update Posted: April 18, 2022
• Sponsor: Institute of Tropical Medicine, Belgium
• Collaborators: University of Gondar; Wollo University; University of York; Maastricht University; University Hospital, Antwerp
• Information provided by (Responsible Party): Institute of Tropical Medicine, Belgium

Study Description

Brief Summary:

Cutaneous leishmaniasis manifestations range from self-healing localized skin ulcers/nodules to diffusely spread chronic lesions. Knowledge on the host-parasite interactions underpinning the different clinical presentations is scarce, in particular for L. aethiopica infections where disease can be extremely severe. Our aim is to define differences in skin immune responses and parasite virulence in CL patients at single cell/parasite level and how it underpins the different clinical presentations (localised, mucocutaneous and diffuse), by producing the first spatially-resolved 'ecological' map of the lesions.

Condition or disease	Intervention/treatment
Cutaneous Leishmaniases	Diagnostic Test: skin biopsy; Diagnostic Test: venous blood sample (plasma, PBMC, WB); Genetic: venous blood sample (HLA); Genetic: skin slit

Detailed Description:

Specific objectives:

1. To profile the full heterogeneity in skin and lesion immunity (single cell RNAseq), and the cellular microenvironment surrounding infected and non-infected macrophages (digital spatial profiling).
2. To study the genomic diversity of L. aethiopica and identify features associated with the different clinical presentations (whole genome sequencing).
3. To understand how parasites respond to the microenvironmental conditions and define parasite survival niches (digital spatial profiling).
4. Study metabolic determinants of skin immunity (e.g. lipid metabolism, bioenergetics, short-chain fatty acids) in the context of key structural features of the skin landscape known to influence local metabolism and immune response (e.g. adipose tissue, follicles, microvasculature) (SpatialOMx).
5. To investigate the association between patient outcomes and the above host/parasite factors at baseline.

Study Design

• Study Type: Observational [Patient Registry]
• Estimated Enrollment: 200 participants
• Observational Model: Case-Control
• Time Perspective: Prospective
• Target Follow-Up Duration: 6 Months
• Official Title: Spatial Analysis of Host-parasite Interactions in the Skin Across the Clinical Spectrum of Cutaneous Leishmaniasis in Ethiopia
• Actual Study Start Date: March 21, 2022
• Estimated Primary Completion Date: September 30, 2023
• Estimated Study Completion Date: December 30, 2023

Groups and Cohorts

Group/Cohort	Intervention/treatment
Local cutaneous leishmaniasis patients group (LCL); local cutaneous leishmaniasis patients	Diagnostic Test: skin biopsy; 4mm skin biopsy; Diagnostic Test: venous blood sample (plasma, PBMC, WB); venous blood sample to acquire plasma, PBMCs and whole blood; Genetic: venous blood sample (HLA); venous blood sample used for HLA typing; Genetic: skin slit; genome sequencing of parasite DNA that is extracted from the skin slit
Mucocutaneous leishmaniasis patients group (MCL); mucocutaneous leishmaniasis patients	Diagnostic Test: skin biopsy; 4mm skin biopsy; Diagnostic Test: venous blood sample (plasma, PBMC, WB); venous blood sample to acquire plasma, PBMCs and whole blood; Genetic: venous blood sample (HLA); venous blood sample used for HLA typing; Genetic: skin slit; genome sequencing of parasite DNA that is extracted from the skin slit
Diffuse cutaneous leishmaniasis patients group (DCL); diffuse cutaneous leishmaniasis patients	Diagnostic Test: skin biopsy; 4mm skin biopsy; Diagnostic Test: venous blood sample (plasma, PBMC, WB); venous blood sample to acquire plasma, PBMCs and whole blood; Genetic: venous blood sample (HLA); venous blood sample used for HLA typing; Genetic: skin slit; genome sequencing of parasite DNA that is extracted from the skin slit
Healthy control patients group Ethiopia (HC - Ethiopia); healthy control patients undergoing elective surgery in Northern Ethiopia	Diagnostic Test: skin biopsy; 4mm skin biopsy
Healthy control patients group Belgium (HC - Belgium); healthy control patients undergoing plastic surgery in Belgium	Diagnostic Test: skin biopsy; 4mm skin biopsy

Outcome Measures

Primary Outcome Measures :

1. Spatially resolved immunological characterization of the CL lesion using single cell RNA sequencing and digital spatial profiling [ Time Frame: Day 0 ]
   Using single cell RNA sequencing and digital spatial profiling methods, we will profile the full heterogeneity in healthy skin/lesion immunity and the cellular microenvironment surrounding infected and non-infected macrophages, respectively.
2. Genomic characterization of L. aethiopica using whole genome sequencing [ Time Frame: Day 0 ]
   Whole genome sequencing will allow us to study the genomic diversity of L. aethiopica and identify features associated with the different clinical presentations.
3. Defining microenvironment and parasite niches in CL lesions using digital spatial profiling [ Time Frame: Day 0 ]
   The digital spatial profiling will indicate the different microenvironmental conditions and parasite survival niches.
4. Spatially resolved determination of the metabolic profile of the CL lesion using spatial OMx [ Time Frame: Day 0 ]
   The metabolic determinants of skin immunity (e.g. lipid metabolism, bioenergetics, short-chain fatty acids) in the context of key structural features of the skin landscape known to influence local metabolism and immune response (e.g. adipose tissue, follicles, microvasculature) will be studied by SpatialOMx.
5. The association between host/parasite factors and patients after treatment using clinical parameters [ Time Frame: Month 6 ]
   Patients are clinically assessed at day 0 (baseline visit), day 28 and month 6. These clinical assessments include a medical questionnaire and lesion assessment, and are compared with the single cell RNA sequencing and spatial resolution data to define potential causal relations between patient outcomes and immunometabolic factors.

Biospecimen Retention:   Samples With DNA

Blood, stool and skin samples will be isolated from CL patients and solely skin samples from healthy controls.

Eligibility Criteria

• Ages Eligible for Study: 12 Years to 40 Years (Child, Adult)
• Sexes Eligible for Study: All
• Sampling Method: Probability Sample

Study Population

Patients presenting at the LRTC and Boru Meda Hospital with clinically suspected CL during the study period will be screened for eligibility. Suspicion for CL is assessed by physicians as per routine care. In general, children are included in the study population since they are commonly affected, and also make up the majority of DCL patients.

Criteria

Inclusion Criteria:

• Willing and able to provide informed consent
• Clinically confirmed CL diagnosis
• Between 12 and 40 years of age

Exclusion Criteria:

• Difficult or too painful sampling zone (see skin biopsy procedure below)
• (Primary) lesion size < 1 cm
• Already receiving CL treatment or received CL treatment in the last 3 months (excluding traditional medicine)
• Known major comorbidity at time of diagnosis (e.g. VL, HIV, TB, malaria, severe intestinal helminth infection)
• Medical history of VL
• Severely underweight (BMI<16)
• Known pregnancy
• Use of immunosuppressive medication in the last month
• Known excessive alcohol use (between >10 intakes/day and >10 intakes/week)
• History of hypersensitivity to local anaesthetics
• Presence of keloids/hypertrophic scars

Contacts and Locations

Contacts

Contact: Thao-Thy Pham, PhD; +32474642614; thaothypham@itg.be

Contact: Wim Adriaensen, PhD; +32 494 75 53 60; wadriaensen@itg.be

Locations

Ethiopia

University of Gondar; Recruiting

Gondar, Amhara, Ethiopia, 6200

Contact: Mikias Woldetensay, MD +251911435963 mikiaswoldetensay@gmail.com

Contact: Silamsaw Asrem Mezgebu, Ms +251921576259 msilamsaw@gmail.com

Sponsors and Collaborators

Institute of Tropical Medicine, Belgium

University of Gondar

Wollo University

University of York

Maastricht University

University Hospital, Antwerp

Investigators

• Principal Investigator: Wim Adriaensen, PhD; Institute of Tropical Medicine Antwerp
• Principal Investigator: Annisa Befekadu Tesfaye, MD; University of Gondar
• Principal Investigator: Seid Getahun, MD; Wollo University
• Principal Investigator: Mikias Woldetensay, MD; University of Gondar

More Information

Additional Information:

Funder - Dioraphte 

• Responsible Party: Institute of Tropical Medicine, Belgium
• ClinicalTrials.gov Identifier: NCT05332093
• Other Study ID Numbers: 1451/20
• First Posted: April 18, 2022
• Last Update Posted: April 18, 2022
• Last Verified: April 2022

Individual Participant Data (IPD) Sharing Statement:

• Plan to Share IPD: Yes
• Plan Description: IPD accessible by managed access
• Supporting Materials: Study Protocol; Statistical Analysis Plan (SAP); Informed Consent Form (ICF)
• Time Frame: After completion of the primary publication
• Access Criteria: Applicants will need to fill out a data access request form
• URL: https://www.itg.be/E/data-sharing-open-access

• Studies a U.S. FDA-regulated Drug Product: No
• Studies a U.S. FDA-regulated Device Product: No

Keywords provided by Institute of Tropical Medicine, Belgium:

leishmania; leishmaniasis; cutaneous leishmaniasis; neglected tropical disease; immunoparasitology

Additional relevant MeSH terms:

Leishmaniasis; Leishmaniasis, Cutaneous; Euglenozoa Infections; Protozoan Infections; Parasitic Diseases; Infections; Skin Diseases, Parasitic; Vector Borne Diseases; Skin Diseases, Infectious; Skin Diseases