HAP1 loss confers l-asparaginase resistance in ALL by downregulating the calpain-1-Bid-caspase-3/12 pathway

Abstract

l-Asparaginase (l-ASNase) is a strategic component of treatment protocols for acute lymphoblastic leukemia (ALL). It causes asparagine deficit, resulting in protein synthesis inhibition and subsequent leukemic cell death and ALL remission. However, patients often relapse because of the development of resistance, but the underlying mechanism of ALL cell resistance to l-asparaginase remains unknown. Through unbiased genome-wide RNA interference screening, we identified huntingtin associated protein 1 (HAP1) as an ALL biomarker for l-asparaginase resistance. Knocking down HAP1 induces l-asparaginase resistance. HAP1 interacts with huntingtin and the intracellular Ca2+ channel, inositol 1,4,5-triphosphate receptor to form a ternary complex that mediates endoplasmic reticulum (ER) Ca2+ release upon stimulation with inositol 1,4,5-triphosphate3. Loss of HAP1 prevents the formation of the ternary complex and thus l-asparaginase-mediated ER Ca2+ release. HAP1 loss also inhibits external Ca2+ entry, blocking an excessive rise in [Ca2+]i, and reduces activation of the Ca2+-dependent calpain-1, Bid, and caspase-3 and caspase-12, leading to reduced number of apoptotic cells. These findings indicate that HAP1 loss prevents l-asparaginase–induced apoptosis through downregulation of the Ca2+-mediated calpain-1-Bid-caspase-3/12 apoptotic pathway. Treatment with BAPTA-AM [1,2-bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid tetrakis(acetoxymethyl ester)] reverses the l-asparaginase apoptotic effect in control cells, supporting a link between l-asparaginase-induced [Ca2+]i increase and apoptotic cell death. Consistent with these findings, ALL patient leukemic cells with lower HAP1 levels showed resistance to l-asparaginase, indicating the clinical relevance of HAP1 loss in the development of l-asparaginase resistance, and pointing to HAP1 as a functional l-asparaginase resistance biomarker that may be used for the design of effective treatment of l-asparaginase-resistant ALL.

Concurrent use of rabacfosadine and L‐asparaginase for relapsed or refractory multicentric lymphoma in dogs

“…Instead, it is possible that L‐ASP might induce metabolic alterations or instill other molecular changes that induce resistance to this protocol. Resistance to L‐ASP in human ALL can be induced by loss of huntingtin‐associated protein 1 . Huntingtin‐associated protein 1 interacts with other proteins to form a complex that mediates Ca 2+ release from the endoplasmic reticulum and through other mechanisms can decrease entry of external Ca 2+ into the cell.…”

Asparagine: A Metabolite to Be Targeted in Cancers

“…Along this line, activation of autophagy by L-asparaginase treatment has been found to protect ALL cells from death, even though it is still unclear whether intracellular asparagine levels can be restored through this mechanism or not [ 64 ] ( Figure 2 ). Another study using genome-wide RNAi screen identified huntingtin associated protein 1 (HAP1) as a biomarker and a driver for the sensitivity to L-asparaginase treatment [ 65 ]. As a result, loss of HAP1 confers resistance to L-asparaginase treatment through preventing the release of calcium from endoplasmic reticulum (ER) and the subsequent calcium-dependent cell death [ 65 ] ( Figure 2 ).…”

“…Another study using genome-wide RNAi screen identified huntingtin associated protein 1 (HAP1) as a biomarker and a driver for the sensitivity to L-asparaginase treatment [ 65 ]. As a result, loss of HAP1 confers resistance to L-asparaginase treatment through preventing the release of calcium from endoplasmic reticulum (ER) and the subsequent calcium-dependent cell death [ 65 ] ( Figure 2 ). Indeed, pharmacogenetic and functional genomic approaches have been used to identified genes associated with L-asparaginase resistance/sensitivity in ALL patient samples or cell lines [ 66 , 67 , 68 , 69 , 70 ].…”

Insights on Metabolic Reprogramming and Its Therapeutic Potential in Acute Leukemia

“…In addition, this study showed that the deprivation of Asn specifically determined a significant induction of zinc finger and BTB domain containing 1 (ZBTB1), which is a transcription factor involved in T cell development [117,118], whose loss was sufficient to sensitize resistant T-ALL cells to ASNase both in vitro and in vivo. ASNase resistance was also recently associated with huntingtin associated protein 1 (HAP1) expression in ALL cells through an experimental approached based on an unbiased genome-wide RNA interference screen [119]. Mechanistically, HAP1 interacts with huntingtin and the intracellular Ca 2+ channel, inositol 1,4,5-triphosphate receptor (IP3R), to form a ternary complex that mediates endoplasmic reticulum (ER) Ca 2+ release upon stimulation with inositol 1,4,5-triphosphate (IP3).…”