Oh, random additional note–have you considered William & Mary?
I am asking because my S24 and a lot of similar kids we know ended up with broadly similar lists to yours, including having Pitt as one of their Likelies. But at a high level, it could be a bit of a struggle finding really suitable Targets in between the Pitt sort of option (which to be fair is a very compelling option for Bio stuff and in general) and then the Reachy schools like WashU and Emory.
In theory some of the Boston schools SHOULD be in that range, but being in Boston they get crazy numbers of applicants, there can be observable anomalies sometimes known as yield protection, and so on.
OK, so at least for the sorts of kids who like Yale, Penn, Brown, WashU, Emory, and so on, some of the universities that actually do fit in that zone are Wake Forest, Rochester, and Case Western. All of which basically are very strong academically, but are not in Boston and so have a lot less unpredictability, and in fact Rochester and Case have merit. So you could consider those.
But personally, my favorite of all the ones that our kids would put in that zone was William & Mary. It has a great campus in a really nice location (like, proximate to beaches and significant cities). The size and format is much more like a medium-sized private than a typical public. The student culture is academicky but quirky and fun. It is relatively undergraduate focused. It has a variety of academic strengths, but Life Sciences were one of them. Even OOS, they are less expensive than full pay private. And then they have a type of Honors program, the Monroe Scholars, with a really nice dorm preference and also special research opportunities.
Of course maybe you have already considered all this and decided against it. I just thought I would mention it as I do think this sort of college (along with the others I mentioned) can help make it feel less risky to not do an ED2.