The dogma "One gene, one protein" is clearly obsolete since cells use alternative splicing and generate multiple transcripts which are translated into protein isoforms, but also use alternative translation initiation sites and termination sites on a given transcripts. Alternative open reading frames for individual transcripts give proteins (the alternative Proteins: AltProts) originate from the 5'- and 3'- UTR mRNA regions, frameshifts of mRNA ORFs or from non-coding RNAs. To gain insight into the role of these newly identified alternative proteins in the regulation of cellular functions, it is crucial to assess their dynamic modulation within a framework of altered physiological modifications such as experimental spinal cord injury (SCI). Here, we carried out a longitudinal proteomic study on rat SCI from 12h to 10 days. Based on AltProt predictions, it was possible to identify a plethora of newly predicted protein hits. Among these proteins, some presented a special interest due to high homology with variable chain regions of immunoglobulins. We focus our interest on the one related to Kappa variable light chains which is similarly highly produced by B-cells in the Bence jones disease, but here expressed in astrocytes. This protein, name Heimdall is an Intrinsically disordered protein which is secreted under inflammatory conditions. We investigated Heimdall's function in these cells using advanced neuroproteomics supported by molecular cell biology and electrophysiology assays and CRISP-CAS9 invalidation. Immunoprecipitation experiments showed that the Heimdall interactome contained proteins related to astrocyte fate keepers such as "NOTCH1, EPHA3, IPO13". However, when Heimdall protein was neutralized utilizing a specific antibody or its gene knocked out by CRISPR-Cas9, sprouting elongations were observed in the corresponding astrocytes. Interestingly, depolarization assays and intracellular calcium measurements in Heimdall KO, established a depolarization effect on astrocyte membranes KO cells impacting the astrocyte phenotype sustained by the decrease of NOTCH2 and switch to neural progenitor cells. Taken together, Heimdall is a novel neural key player involved in astrocytes gatekeeper phenotype.
References
Capuz A, et al., Heimdall, an alternative protein issued from a ncRNA related to kappa light chain variable region of immunoglobulins from astrocytes: a new player in neural proteome, Cell Death Dis. 2023 Aug 16;14(8):526
Capuz A, et al., Astrocytes express aberrant immunoglobulins as putative gatekeeper of astrocytes to neuronal progenitor conversion. Cell Death Dis. 2023 Apr 4;14(4):237.
Capuz et al., The Antibody Dependant Neurite Outgrowth Modulation Response Involvement in Spinal Cord Injury. Front Immunol. 2022 Jun 16;13:882830.