Tetraspanin-induced death of myeloma cell lines is autophagic and involves increased UPR signalling.
BACKGROUND
Multiple myeloma (MM) therapy is hindered by the interaction of the heterogeneous malignant plasma cells with their microenvironment and evolving drug resistance. We have previously shown that the membranal tetraspanins, CD81 and CD82, are under-expressed in MM cells and that their reintroduction causes massive non-apoptotic death. In this study, we aimed to characterise the tetraspanin-induced MM death.


METHODS
Multiple myeloma cell lines were transiently transfected with eGFP-CD81N1/CD82N1 fusion proteins and assessed for death mode by flow cytometry (propidium iodide, ZVAD-fmk, 3MA), activation of unfolded protein response (UPR), and autophagy (immunoblot, RT-PCR).


RESULTS
Cell death induced by CD81N1 and CD82N1 in MM cell lines was autophagic and involved endoplasmic reticulum (ER)-stress manifested by activation of UPR pathways, PERK (protein kinase-like ER kinase) and IRE1 (inositol-requiring 1). We also established the relative X-box binding protein 1 baseline expression levels in a panel of MM cell lines and their general dependence on autophagy for survival. Timeline of UPR cascades and cell fate supported our results.


CONCLUSIONS
This is the first publication implicating tetraspanins in UPR signalling pathways, autophagy, and autophagic death. Integration of our findings with published data highlights the unifying dependence of MM cells on ER-Golgi homoeostasis, and underscores the potential of tetraspanin complexes and ER-stress as leverage for MM therapy.
Zismanov
V
V
Meir Medical Center, Kfar Saba, Israel.
Lishner
M
M
Tartakover-Matalon
S
S
Radnay
J
J
Shapiro
H
H
Drucker
L
L
D015703:Antigens, CD / Q000502:physiology*; D017209:Apoptosis; D001343:Autophagy; D045744:Cell Line, Tumor; D002470:Cell Survival; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D006801:Humans; D051861:Kangai-1 Protein / Q000502:physiology*; D009101:Multiple Myeloma / Q000473:pathology* / Q000628:therapy; D017510:Protein Folding; D015398:Signal Transduction; D060225:Tetraspanin 28
D015703:Antigens, CD; C554798:CD81 protein, human; C497688:CD82 protein, human; D051861:Kangai-1 Protein; D060225:Tetraspanin 28