Immunohistochemical Detection of the Chaperone-Mediated Autophagy Markers LAMP2A and HSPA8 in Formalin-Fixed and Paraffin-Embedded Tissues

Tereza Losmanová; Mario P Tschan; José A Galván; Sabina Berezowska

doi:10.1007/978-1-0716-2675-7_11

Immunohistochemical Detection of the Chaperone-Mediated Autophagy Markers LAMP2A and HSPA8 in Formalin-Fixed and Paraffin-Embedded Tissues

Methods Mol Biol. 2023:2566:141-147. doi: 10.1007/978-1-0716-2675-7_11.

Authors

Tereza Losmanová¹, Mario P Tschan¹, José A Galván¹, Sabina Berezowska^{2

3}

Affiliations

¹ Institute of Pathology, University of Bern, Bern, Switzerland.
² Institute of Pathology, University of Bern, Bern, Switzerland. [email protected].
³ Department of Laboratory Medicine and Pathology, Institute of Pathology, Lausanne University Hospital and University of Lausanne, Lausanne, Switzerland. [email protected].

PMID: 36152248
DOI: 10.1007/978-1-0716-2675-7_11

Abstract

Autophagy is crucial for maintaining cellular homeostasis and its deregulation is involved in disease development, including cancer. The key players of chaperone-mediated autophagy (CMA), a particular selective subtype of autophagy, are HSPA8 and LAMP2A. Both proteins can be immunohistochemically detected in formalin-fixed paraffin-embedded (FFPE) tissue. LAMP2A is frequently overexpressed in a variety of cancers where it likely supports cancer cell survival and resistance to anti-cancer therapies in a context-dependent manner. Here we present the immunohistochemical staining protocol of antibodies against LAMP2A and HSPA8, using an automated staining system, suitable for routine diagnostics. Additionally, we also suggest a staining evaluation method.

Keywords: CMA; Chaperone-mediated autophagy; HSPA8; Immunohistochemistry; LAMP2A.

MeSH terms

Autophagy / physiology
Chaperone-Mediated Autophagy*
Formaldehyde / metabolism
Lysosomal-Associated Membrane Protein 2 / metabolism
Lysosomes / metabolism
Molecular Chaperones / metabolism
Paraffin Embedding

Substances

Lysosomal-Associated Membrane Protein 2
Molecular Chaperones
Formaldehyde