Single-cell spatial profiling reveals immune-steroidogenic crosstalk in adrenals of patients with primary aldosteronism

Noorzaileen Eileena Zaidi; Amnani Aminuddin; Aina Nadheera Abd Rahman; Faeezah Abdul Latif; Emily Goodchild; Kate Laycock; Eva Wozniak; Charles Mein; Muaatamarulain Mustangin; Nor Adzimah Johdi; Nor Haslinda Abd Aziz; Adli Ali; Azraai Bahari Nasruddin; Miroslav Solar; Troy Puar Hai Kiat; Norlela Sukor; William Drake; Morris Jonathan Brown; Elena Aisha Azizan

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Single-cell spatial profiling reveals immune-steroidogenic crosstalk in adrenals of patients with primary aldosteronism

Author: Noorzaileen Eileena Zaidi ¹ ; Amnani Aminuddin ¹ ; Aina Nadheera Abd Rahman ¹ ; Faeezah Abdul Latif ¹ ; Emily Goodchild ² ; Kate Laycock ² ; Eva Wozniak ³ ; Charles Mein ³ ; Muaatamarulain Mustangin ⁴ ; Nor Adzimah Johdi ⁵ ; Nor Haslinda Abd Aziz ⁶ ; Adli Ali ⁷ ; Azraai Bahari Nasruddin ⁸ ; Miroslav Solar ⁹ ; Troy Puar Hai Kiat ¹⁰ ; Norlela Sukor ¹¹ ; William Drake ¹² ; Morris Jonathan Brown ¹³ ; Elena Aisha Azizan ¹⁴
Author Information

1. Department of Medicine, Faculty of Medicine, The National University of Malaysia, Malaysia
2. Centre of Clinical Pharmacology and Precision Medicine, William Harvey Research Institute, Queen Mary University of London, London, United Kingdom
3. Barts and London Genome Centre, School of Medicine and Dentistry, Blizard Institute, London, United Kingdom
4. Department of Pathology, Faculty of Medicine, The National University of Malaysia, Malaysia
5. UKM Medical Molecular Biology Institute, The National University of Malaysia, Malaysia
6. Department of Obstetrics and Gynaecology, Faculty of Medicine, The National University of Malaysia, Malaysia
7. Department of Paediatrics, Faculty of Medicine, The National University of Malaysia, Malaysia; Research Center, Hospital Tunku Ampuan Besar Tuanku Aishah Rohani, Universiti Kebangsaan Malaysia (UKM) Specialist Children’s Hospital, Malaysia
8. Department of Endocrinology, Hospital Putrajaya, Malaysia
9. 1st Department of Internal Medicine–Cardioangiology, Charles University Faculty of Medicine in Hradec Kralove and University Hospital Hradec Kralove, Czech Republic
10. Department of Endocrinology, Changi General Hospital, Singapore
11. Department of Medicine, Faculty of Medicine, The National University of Malaysia, Malaysia; Hospital Canselor Tuanku Muhriz Universiti Kebangsaan Malaysia, Malaysia
12. NIHR Barts Biomedical Research Centre, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London, United Kingdom; St Bartholomew’s Hospital, Barts Health NHS Trust, London, United Kingdom
13. Centre of Clinical Pharmacology and Precision Medicine, William Harvey Research Institute, Queen Mary University of London, London, United Kingdom; NIHR Barts Biomedical Research Centre, Barts and The London School of Medicine and Dentistry, Queen Mary University of London, London, United Kingdom
14. Department of Medicine, Faculty of Medicine, The National University of Malaysia, Malaysia; entre of Clinical Pharmacology and Precision Medicine, William Harvey Research Institute, Queen Mary University of London, London, United Kingdom; Research Center, Hospital Tunku Ampuan Besar Tuanku Aishah Rohani, Universiti Kebangsaan Malaysia (UKM) Specialist Children’s Hospital, Malaysia
Publication Type:Journal Article
MeSH: Hyperaldosteronism; Humans
From: Journal of the ASEAN Federation of Endocrine Societies 2026;41(S1):6-
CountryPhilippines
Language:English
Abstract: Introduction:Primary aldosteronism (PA), most commonly caused by aldosterone-producing adenomas (APAs), represents the leading form of surgically curable secondary hypertension. While genomic studies have elucidated the mutational landscape of APAs, the spatial organisation and functional role of immune populations across APAs, aldosterone-producing micronodules (APMs), and adjacent adrenal cortex remain poorly defined at single-cell resolution.
Methodology:Single-cell RNA sequencing (scRNA-seq) was integrated with spatial transcriptomics in APAs and paired adjacent adrenal cortex, complemented by immunohistochemical (IHC) validation. Immune populations were spatially mapped using canonical markers (CD14, CD68, CD163, HLA-DR, CD8A, and CD4) across defined adrenocortical regions.
Results:The adrenal microenvironment in PA demonstrates structured immune organisation rather than passive infiltration. CD14+ monocyte-lineage cells localize intraparenchymally within APAs (n = 10), intercalating between CYP11B2+ aldosterone-producing cells and forming a pattern distinct from perivascular immune niches. scRNA-seq further identified a transcriptionally distinct CD14+ population within the zona reticularis (zR) that co-expresses steroidogenic markers (CYB5A, SULT2A1, TSPAN12) while lacking canonical monocyte transcripts. IHC supported this observation, demonstrating CD14 expression within adrenocortical zR parenchymal cells (n = 5). In parallel, CD4 and HLA-DRA exhibited diffuse cytoplasmic staining within zR parenchymal cells in the absence of classical macrophage marker coexpression (CD68), suggesting non-canonical or context-dependent expression within steroidogenic compartments. CD68+ and CD163+ macrophages were sparsely distributed across APA, APM and adjacent cortex, consistent with lowdensity tissue-resident populations, while CD8A+ cytotoxic lymphocytes were enriched in APAs and APMs with diffuse parenchymal cytoplasmic staining of CD8A additionally observed within the zR.
Conclusion:These findings reveal a previously unrecognized spatially organised immune-steroidogenic interface within the adrenal cortex. The presence of immune-associated transcriptional and protein signatures within zR cells suggests potential functional plasticity of steroidogenic cells, possibly extending to antigen presentation-related pathways, warranting further mechanistic investigation
Full text:202607280929466407OP_A006.pdf