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

Spatial Analysis of Host-parasite Interactions in the Skin Across the Clinical Spectrum of Cutaneous Leishmaniasis in Ethiopia

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.

Study Overview

• Status: Active, not recruiting
• Conditions: Cutaneous Leishmaniases
• Intervention / Treatment: 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 Type: Observational
• Enrollment (Actual): 92

Contacts and Locations

• Study Contact: Name: Thao-Thy Pham, PhD; Phone Number: +32474642614; Email: thaothypham@itg.be
• Study Contact Backup: Name: Wim Adriaensen, PhD; Phone Number: +32 494 75 53 60; Email: wadriaensen@itg.be

Study Locations

• Ethiopia
  • Amhara
    • Gondar, Amhara, Ethiopia, 6200
      • University of Gondar

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 12 years to 40 years (Child, Adult)
• Accepts Healthy Volunteers: N/A

• 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.

Description

Inclusion Criteria:

• Willing and able to provide informed consent
• Clinically confirmed CL diagnosis
• Between 12 and 50 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

Study Plan

How is the study designed?

Design Details

• Observational Models: Case-Control
• Time Perspectives: Prospective

Number of groups / cohorts

5

Cohorts and Interventions

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

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Spatially resolved immunological characterization of the CL lesion using single cell RNA sequencing and digital spatial profiling	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.	Day 0
Genomic characterization of L. aethiopica using whole genome sequencing	Whole genome sequencing will allow us to study the genomic diversity of L. aethiopica and identify features associated with the different clinical presentations.	Day 0
Defining microenvironment and parasite niches in CL lesions using digital spatial profiling	The digital spatial profiling will indicate the different microenvironmental conditions and parasite survival niches.	Day 0
Spatially resolved determination of the metabolic profile of the CL lesion using spatial OMx	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.	Day 0
The association between host/parasite factors and patients after treatment using clinical parameters	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.	Month 6

Collaborators and Investigators

• Sponsor: Institute of Tropical Medicine, Belgium
• Collaborators: University Hospital, Antwerp; Maastricht University; University of York; University of Gondar
• Investigators: Principal Investigator: Wim Adriaensen, PhD, Institute of Tropical Medicine Antwerp; Principal Investigator: Mikias Woldetensay, MD, University of Gondar

Publications and helpful links

Helpful Links

• Funder - Dioraphte

Study record dates

Study Major Dates

• Study Start (Actual): March 21, 2022
• Primary Completion (Actual): March 30, 2024
• Study Completion (Estimated): December 31, 2024

Study Registration Dates

• First Submitted: March 8, 2022
• First Submitted That Met QC Criteria: April 11, 2022
• First Posted (Actual): April 18, 2022

Study Record Updates

• Last Update Posted (Actual): April 17, 2024
• Last Update Submitted That Met QC Criteria: April 16, 2024
• Last Verified: April 1, 2024

More Information

Terms related to this study

• Keywords: leishmaniasis; leishmania; cutaneous leishmaniasis; neglected tropical disease; immunoparasitology
• Additional Relevant MeSH Terms: Skin Diseases; Infections; Vector Borne Diseases; Parasitic Diseases; Protozoan Infections; Skin Diseases, Parasitic; Skin Diseases, Infectious; Euglenozoa Infections; Leishmaniasis; Leishmaniasis, Cutaneous
• Other Study ID Numbers: 1451/20

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: YES
• IPD Plan Description: IPD accessible by managed access
• IPD Sharing Time Frame: After completion of the primary publication
• IPD Sharing Access Criteria: Applicants will need to fill out a data access request form
• IPD Sharing Supporting Information Type: STUDY_PROTOCOL; SAP; ICF

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No