KCTD15 is overexpressed in human childhood B-cell acute lymphoid leukemia

Smaldone et al., Scientific Reports, 2019 went looking for new molecular players in childhood B-cell acute lymphoblastic leukaemia by sequencing leukaemic cells from three children against normal B cells from three healthy adults. Among the 25 most strongly upregulated genes, 11 had never been connected to leukaemia at all. The authors picked KCTD15, a member of a protein family better known from brain disease and obesity genetics, and confirmed its overexpression in a second group of 12 patients and in four B-cell leukaemia lines. In patients the transcript fell 3.6 fold between diagnosis and day 33 after induction therapy, and the protein fell with it.

To test whether the gene was doing anything or merely marking the disease, the authors silenced it in RS4;11, a leukaemia line carrying the MLL rearrangement. They used self-delivering antisense oligonucleotides added straight to the culture medium, with a scrambled oligonucleotide and an untreated arm as controls, choosing 8 μM after testing 4 and 8. Uptake measured with a fluorescently labelled control oligonucleotide was about 99.2%, assessed after washing surface-bound material away, and no transfection reagent appears anywhere in the methods. Because the deepest silencing in this line needed a long incubation, the experiment ran to 16 days with fresh oligonucleotide added whenever the cells were split.

The result was clean. Transcript and protein both fell progressively from day 8 to day 16, and the proportion of dead cells rose from 26.3% to about 80%, while the scrambled control produced no appreciable increase over untreated cells. One caution governs how this paper may be used: it names no supplier for the oligonucleotides, identifying them only by chemistry and citing published methodology papers. The product named on this record is established from AUM's own records rather than from anything the paper says.