Question: So, spherical mineral everywhere, with concentric growth rings - the organics in the ring usually contain osteopontin as we have shown in several cases. Have you evidence for that in your deposits?

to the audience: is the ciliary body calcification contribute to glaucoma?

have you seen changes in general mitochondrial shape?

It is striking that in vitro the calcifications start intracellular, since PPi is low extracellularly in ODD. Could it be that the extracellular calcifications in ODD patients originate from Bruch's membrane?

Question: Is it possible that all drusen are in fact calcified as hard drusen, and that acidic sample preparations has led to the term soft drusen? If that is true, maybe calcification is the initiating event.

Have you checked the molecular markers of the cells at the end of the culture period, in case they have transdifferentiated?

Question: Unlike connective tissue cells, it is both remarkable and unusual that epithelial cells in general can produce mineralized nodules. So, like in vivo, have you checked if your RPE cells make collagen and elastin as is found in Bruch's Membrane and supposedly made by RPE cells?

If understand well, you are doing really long term experiments with these primary porcine RPE cells. Isn't scenescence a contributor/issue in the HA sphere development? Did you tested for scenescence markers? maybe your small molecular screen rather is an anti senolytics screen?

Do you condition the culture medium with inorganic phosphate to accelerate calcification?

AR signaling is also important in vasculature development in females. Do you plan to also study the VSMC in response to testosterone in female mice? Would be interesting to see that to...

Does the calcifying effect are only due to testosterone or the balance between oestrogens and testosterone ???

any idea why the thalamus?

Imre, how can you be sure that Os680 in the vessel is an artefact while that of the Bruch membrane is specific? Do you have additional evidence? What does the 1 penny AR and Von Kossa staining show?

is there any phosphorylation with Sortilin that could be blocked as a treatment ?

Oliver, I sent you a note

Is it possible that the PXE patients also have higher calcifications in the trachea ?

Do you have an idea what causes the tortuosity of the epithelia? Is it also present in patients?

olivier : 1) Would you sufficiently relay on PPi dosage for the diagnosis ? and 2) would you treat these patients with Low PPi ???

Is it well accepted that the main site of ABCC6 expression is mainly liver/kidney, or are there possibilities of expression in other tissues?

In animal models it has already been tested, but not with therapeutic objective.

I am wondering if the presenters can comment on the possibility of gene replacement therapy for PXE by over expressing ABCC6 in the liver/kidney

What makes these iPSC fibroblasts lung fibroblast-like? Culturing conditions? What are their advantages over skin fibroblasts?

Are there mouse models of systemic sclerosis?

What is the natural history of systemic sclerosis? Is it useful to prevent calcinosis from a clinical standpoint in systemic sclerosis or shouldn't we look for a treatment which resolves existing calcifications in systemic sclerosis patients?

could it be that low PPi is caused by renal failure in the mouse model?