Table 1.

– *+ indicates the timepoint where the named clones were detected.

	APC	TP53	KRAS	NRAS	BRAF	EGFR
Patient 1
Tissue NGS	+	+
ctDNA – T1*	+	+	+		+	+
ctDNA – T2	+	+	+			+
ctDNA – T3	+	+	+		+	+
Patient 2
Tissue NGS	+	+
ctDNA – T1*	+	+
ctDNA – T2	+	+
ctDNA – T3
ctDNA – T4	+	+	+
ctDNA – T5
Patient 3
Tissue NGS	+	+
ctDNA – T1*	+	+
ctDNA – T2*	+	+			+	+
ctDNA – T3	+	+	+			+
ctDNA – T4	+		+			+
ctDNA – T5	+	+	+			+
Patient 4
Tissue NGS	+	+
ctDNA – T1*	+	+	+
ctDNA – T2	+	+	+			+
ctDNA – T3	+	+	+		+	+
ctDNA – T4	+	+	+	+	+
Patient 5
Tissue NGS	+	+
ctDNA – T1*	+	+				+
ctDNA – T2	+	+				+
ctDNA – T3	+	+				+
Patient 6
Tissue NGS	+	+
ctDNA – T1*	+	+	+		+
ctDNA – T2	+	+	+		+
ctDNA – T3	+	+	+		+

Known acquired mechanisms of resistance were noted in all the cases. The sub-clones were noted to be polyclonal, and the number of clones varied over time. Some clones disappeared over time during non-EGFR-based therapy (EGFR holiday such as KRAS clones in patient 2, BRAF clones in patient 3, and EGFR clones in patient 4.