1) there is no concrete evidence to suggest ctDNA to have its origin in CTCs.

ctDNA can be present in

patients who have non detectable levels of CTCs in their blood streams.

2) HER2 receptor is a different receptor than EGFR (HER1) which is found mutated in NSC lung cancer
patients. This is regulated by EGF like ligands, neuregulins etc to create HER1 ,2,3,4 heterodimers.

3) HER2 negative status does not necessarily denote non-amplification of the gene. There are other
methods of regulation whereby HER2 receptors could be downregulated on the cell surface. Hence
you do need an IHC confirmation of the HER2 status of a population of ctc and not single CTC. There
will never be complete absence of HER2 expression. But there will be a mix of two populations and
what needs to be known is what is the proportion of cells that are HER2 positive.

4) HER2 gene amplifaction is only one of the ways of increasing surface expression. There are other
methods as well by which cells can up regulate HER2 on the cell surface.