[spligak]

You're correct about CRISPR Cas9. The off-target affects are difficult to manage.

The paper describes Cas12a2. This is a different mechanism with discovery origins in - of all things - agriculture. It does not attempt in any way to reprogram cells. It uses a guide protein to locate a specific mutation with exacting precision and, when it activates, unleashes total destruction of the cell.

The implications of Cas12a2 on undruggable conditions that exhibit known driver mutation profiles is profound.

Source: I have personally funded novel research based on Cas12a2 for an undruggable condition I have. I have personally seen my condition "cured" in vitro using this technology and it left all of my WT cells unharmed. Some of the researchers I've funded are co-authors in the paper linked. I am a layperson in this field (I'm a SWE, not in biotech), but I am happy to answer questions.

[Gethsemane]

Have you written about your experience anywhere? It would be interesting to see how you approached the research sector as a layperson. Are there any plans to move to in vivo? Best of luck with your research!

[spligak]

> Are there any plans to move to in vivo?

Yes! That's the next step. There wasn't a mouse model for my variant so they're building that, too. But in vivo testing should be underway this calendar year.

[spligak]

I haven't written about it publicly, but I can elaborate here. I don't mind answering further questions about it even if you believe they'd make me uncomfortable - they won't.

I've come to terms with what's happening to my body and that I may not benefit from my efforts.

Background: ~3 years ago I was diagnosed with a very rare MPLW515L-driven blood cancer known as a myeloproliferative neoplasm. My hematopoietic stem cells (HSCs) acquired this mutation and they produce busted downstream products.

Most notably, one of those downstream products are hyper-lobulated megakaryocytes that spew inflammatory cytokines into my bone marrow and destroy the bone marrow niche over time. The destruction happens specifically because the inflammation mobilizes stromal cells and they erroneously produce scar tissue (fibrosis) all along the walls of the good, spongy marrow. There are other sources of damage but this is the one path most aligned to abbreviated survival and transformation into AML.

In effect, my bone marrow is rusting and very slowly failing. The failure could speed up with the acquisition of additional mutations or any other systemic inflammatory condition.

Anyway, 3 years ago my first retail hematologist told me "it's rare, you're fine, take aspirin and go home."

I couldn't accept that - this seemed bad. I decided that if I wanted to know the truth I needed to physically stand in front of the foremost expert in the world on the topic and ask them "what is the state-of-the-art?"

I came to this conclusion after about a year of reading all the most well-cited academic papers about AML, Myelofibrosis, and Essential Thrombocythemia. In particular, anything that mentioned MPL. There are virtually no papers mentioning MPL.

To put that in perspective: 500,000 patients in the US deal with the broad disease category. 5% of those are MPL, and 40% of those are the -K variant. So 10,000 people - which means anything targeting it would be well into orphan drug designation territory. I'd need to find a pretty niche researcher.

So, I laddered up the academic food chain using a little cash (donations), emails, airline tickets, and conference admission. ~2 years after my diagnosis I found myself in a closed-door session called the MPN Roundtable in Chicago with 100 of the foremost experts in the world. No cameras, no transcripts, just some of the greatest minds in the field earnestly debating the path forward to a cure.

I listen carefully to them, ask dumb questions, connect dots across research. I rehomed my care to an academic research hospital specializing in MPN research, and started funding research on the condition it includes my specific MPL mutation. Researchers happily oblige.

Cas12a2 was the keynote topic at this year's meeting and there was _very little_ dissent.

[reenorap]

My aunt had the same disease you mention and was on medications since the 90s. She lead a healthy life with no real side effects from her medication and she passed away last year in her 80s. To be perfectly honest, she did die of the disease, because her medication stopped working and her bone marrow was all scarred. But up until a year before she passed away she was very active and healthy. Once the medication stopped working, she went steadily downhill until she passed away.

Hopefully you get great progress on your research but I just wanted to reassure you that the name sounds scary but the current treatment appears to work well and hopefully gives you enough runway to find your cure.

[smallnix]

> I just wanted to reassure you that the name sounds scary but the current treatment appears to work well

Even a charitable read is condescending. This person just wrote they are in the innermost research circles. I think they are beyond the "scary name" - needs reassurance phase.

[btam]

This is so impressive - kudos to you. Thanks for sharing and being open to questions.

How much overall has this costed you? Do you think that a middle-class person could afford to do what you did?

> So, I laddered up the academic food chain using a little cash (donations), emails, airline tickets, and conference admission. ~2 years after my diagnosis I found myself in a closed-door session called the MPN Roundtable in Chicago with 100 of the foremost experts in the world. No cameras, no transcripts, just some of the greatest minds in the field earnestly debating the path forward to a cure.

Why don't they allow recordings at the MPN Roundtable? It could be useful for others to learn from.

[spligak]

> How much overall has this costed you?

For this project, seed capital low six figures. I am collaborating with family and friends, non-profits, and using doubling mechanisms available to me to at work to fund the very early speculative bench research. This is where we are and its sustainable today.

Once we have the basic tech worth scaling up - to raise the first round of capital, I estimate $1-2m with a wider friends & family and angel investor round. This will be early de-risking research and delivery mechanism testing.

Beyond that I can see a path to a ~$20m round to further de-risk any assets that come out of these speculative efforts, but I haven't gotten this far.

In rare disease therapeutics the challenge isn't raw capital, it's finding the _right_ capital that understands how assets like these get de-risked and can tolerate the shape of the upside. Anything CRISPR-based is usually not a chronic therapeutic, so that disqualifies most of big pharma. Acute, curative technology like this requires informed capital.

> Do you think that a middle-class person could afford to do what you did?

Yes.

In the rare disease field even a small amount of capital attracts enough attention to have meaningful conversation with bench researchers. If you're willing to travel to the niche conferences, ask dumb questions, grind out the studying, and approach the speakers after their sessions.

Researchers respect people who do their homework and mobilize to meet them. They want (need) to hear from patients and caregivers - so they tend to listen very carefully.

Fun fact: I've had multiple researchers ask me for samples of my bone marrow. Has only happened in-person :)

> Why don't they allow recordings at the MPN Roundtable? It could be useful for others to learn from.

I don't know, it's been going on for a while. I can speculate: they're discussing pre-publication data, some of which had come out of their labs only hours prior to their presentation. It's completely unfiltered. I think there's real risk of some of the things that are shown being sensationalized or taken out of context.

The audience is trained and practiced in keeping a sober, skeptical lens on everything they see - so it's more about the debate and tear-down of the early data for the betterment of the niche.

There's zero attempt to hide anything, it's just a forum for collegial debate.

[p1esk]

Do you feel that recent advances in AI can speed up such rare disease research?

[darkwizard42]

Incredible story, just pure resourcefulness and grit in following this through. I know it sucks to have this disease, but kudos for how you approached this.

[andy_ppp]

"When have you most successfully hacked a non-computer system to your advantage?" Amazing resourcefulness, you should consider applying to YC if you haven't! And I hope you manage to find a solution to your problem it sounds very promising.

[CamperBob2]

And by the way, when Anthropic (sic) tells you that it's too dangerous to allow GPT-2/GPT-3/GPT-4/GPT-5/Sonnet/Opus/Mythos/Fable to discuss human biology, and some of us object vociferously to their premise, this is what we're talking about.

[gleenn]

I haven't heard anyone specifically state their justification for blocking bio research along I can only assume it's to prevent manufacturing bio weapons or virii?

[junon]

This is amazing. Thank you for sharing.

[ticulatedspline]

wow, very interesting I can't say I've really ever heard of anyone financing research themselves, hope things work out and maybe a treatment arrives in time for you.

As an aside if you end up cryogenically freezing yourself for a future treatment don't forget to actually cure your boneitis when they thaw you out.

[ordinaryradical]

Seconding this comment. I would love to read a write-up about your experience and how you’ve been trying to work on solutions for yourself. Stories like these are valuable to the field and inspiring to other folks dealing with a tough diagnosis.

[spligak]

Thank you. I want to make sure you see the comment above - I think it was your comment that nudged me into writing it :) Happy to answer any other questions, here or over email (in my user profile).

> other folks dealing with a tough diagnosis

The toughest part has been the spiritual journey. Loneliness unlike anything I've experienced. I felt forgotten without the opportunity to be known in the first place. I was happy - and emotional - to learn I wasn't alone. It took me 2 years, but I've found my people.

[p1esk]

I’d also like to read about your experience.

[spligak]

Done! Wrote a bit more in a comment above. Thank you for the interest.

[acomjean]

We did whole genome crispr designs at my last university job. Can confirm that off target effects are an issue with cas9. Pattern matching across the genome to see if a design is unique takes some time. These were interesting pipelines to work on.

It’s only a matter of time before the next better thing shows up.

[GaggiX]

I know nothing about this field, but I imagine the actual problem is how do you deliver the Cas12a2 protein to each individual cancer cell compare to a viral gene therapy?

[spligak]

There are two major problems, delivery is one of them. Collateral damage of mass cell destruction leading to systemic inflammation is the other.

The approach I'm reviewing now uses lipid nanoparticles (LNPs) for delivery. It isn't great for targeting my bone marrow condition but its workable. The team hasn't optimized it at all, either. There are also viral delivery mechanisms that I haven't studied yet.

The collateral damage problem is the backpressure on the delivery problem. If you get really good at delivery, you can destroy A LOT of cells very quickly. The human body (usually) responds to these events by releasing a lot of pro-inflammatory cytokines. This can lead to cytokine storms or worse.

As you "get good" at killing the target cells, the net effect can turn bad. It will probably be a balancing act.

[shevy-java]

Lipid nanoparticles are quite old as-is. How do you target cells specifically?

> If you get really good at delivery, you can destroy A LOT of cells very quickly.

You can destroy cells quickly. Ok. So the question is: how do you detect specifically only cancer cells via lipid nanoparticles? That was already a problem years ago with Herceptin. The rationale that is always used is that "we need to do something" for certain aggressive cancers. It has never been a super-effective technique, despite all the promo of how monoclonal antibodies are so accurate.

> As you "get good" at killing the target cells, the net effect can turn bad. It will probably be a balancing act.

That's already the status quo in the whole cancer field. I don't think that more than sloppy accuracy is acceptable for any gene therapy - and the off-target cleaving of CRISPR has always been the number #1 problem here.

[spligak]

> I don't think that more than sloppy accuracy is acceptable for any gene therapy

Valid critiques of Cas12a2 must acknowledge the mechanistic differences between Cas9 and Cas12a2. There is no research to suggest Cas12a2 is "sloppy" and significant research that demonstrates it is not "sloppy."

I appreciate the skepticism but I would encourage you to study the actual mechanism discussed in the paper.

[cyberax]

> So the question is: how do you detect specifically only cancer cells via lipid nanoparticles?

You don't. Healthy cells will also get these nanoparticles, but without the triggering DNA sequence, the mRNA payload will remain inert and eventually will be degraded.

[spligak]

> Healthy cells will also get these nanoparticles, but without the triggering DNA sequence, the mRNA payload will remain inert and eventually will be degraded.

This is my understanding as well.

[im3w1l]

Naively, I would deal with this by deciding how many cells I want to kill each day and then figure out a dosing schedule that achieves that. Or maybe it's better to do one dose every few days. But yeah either way.

[spligak]

> I would deal with this by deciding how many cells I want to kill each day

Yes, this is an approach. This starts to exceed my understanding of the problems the teams are facing - but there are concerns about the efficacy of Cas12a2-based approaches fading. Not because the cells adapt, but because your immune system starts acting funny in the presence of the payload.

I don't recall the specifics but there seems to be a window of opportunity for these things.

[shevy-java]

So how does Cas12a2 mitigate off-target effects?

If it were to work, gene therapy as-is would be possible. Which it is not, not even for those overpriced therapies. I have no doubt that sooner or later it will happen, as the problem space is finite, not infinite, but I simply don't see the correlation here.

> The implications of Cas12a2 on undruggable conditions that exhibit known driver mutation profiles is profound.

So what does this change exactly? Humans defined it as "undruggable conditions". You can reason this is an improvement, but I still see it in failure-territory. If it were to work, gene therapy would be an accurate - and affordable - technique. Which it is not right now.

> I am a layperson in this field (I'm a SWE, not in biotech), but I am happy to answer questions.

How does "answering questions" offset the technology being inferior right now?

[spligak]

> So how does Cas12a2 mitigate off-target effects?

Others in this thread may be able to give a better analogy, but I'll try:

Cas9 is like open heart surgery on millions of cells all at once. We know the specific outcome we want - a surgical replacement of a sliver of a sequence - but just like open heart surgery, it's an inexact operation. Cas9 tolerates mismatches which categorically allows off-target matching. It also operates on DNA, so any off-target effects reprogram the cell's primary source code.

We want the Cas9 "patient" cell to survive.

In contrast, Cas12a2 is key-locked self-destruction switch. It targets single-stranded RNA transcripts with a specific guide protein. So the specificity is two-fold: the guide protein doesn't tolerate mismatches, and its operating on a _downstream byproduct_ of the DNA. When the key (guide protein) matches, it unleashes total destruction within the cell.

We want the Cas12a2 "patient" cell to die.

> If it were to work, gene therapy would be an accurate - and affordable - technique. Which it is not right now.

Correct on the first point. If it were to work, gene therapy could be more common. I do not know how to make it affordable, yet. In the models I've built to commercialize this I estimate a Cas12a2 treatment would cost approximately as much as a bone marrow transplant.

> How does "answering questions" offset the technology being inferior right now?

In fairness, asking and seeking answers to questions is all I have right now. There is no cure to my disease so the upside - no matter how futile you may perceive it to be - to me, is infinite.

If I can solve it I may get a few more years with my daughter. If I can't, I can show her how to live fighting for an answer that may never come.

You're not wrong, you and I just have different perspectives on the upside.

[rozap]

This is wild, have you written about it publicly, or can you expand on it here?

[spligak]

I have now :) I replied to a sibling comment. Thank you for the nudge to share.

[manmal]

May I ask, how much money you have spent so far on this effort?