To our immune system, a potentially lifesaving gene therapy can look a lot like a dangerous infection. That's because most genetic medicine uses viruses or double-stranded DNA to deliver genetic information to target cells. DNA in its traditional double helix form can lead to toxic immune stimulation and be difficult to package into cellular delivery vehicles. As a result, the reach of genetic medicine is limited today.
Kano Therapeutics is taking a different approach to genetic therapies. The company is developing gene-editing technologies using circular single-stranded DNA (cssDNA), a biomolecule that is less toxic than double stranded DNA and more stable than RNA, and could be delivered more efficiently to many parts of the body to treat genetic diseases, cancers, and more.
The company, which was founded by former MIT postdoc Floris Engelhardt, professor of biological engineering Mark Bathe, and John Vroom MBA '22, is developing a platform for manufacturing cssDNA of customized lengths and sequences, which could deliver genetic material to fix or replace faulty genes.
"We can work with CRISPR and other gene-editing technologies," Engelhardt says. "CRISPR finds a location in a genome, binds to it, and cuts at that location. That allows you to edit a gene or stop a gene from functioning. But what if you have a loss-of-function disease where you need to insert a new piece of genetic code? Our approach allows you to replace whole genes or add genetic information."
Making DNA flexible
Around 2019, Bathe's lab published research describing ways to engineer the sequence and length of cssDNA molecules, which have been used in labs for decades but have increasingly drawn interest for improving gene therapies. Several pharmaceutical companies immediately reached out.
"Single-stranded DNA is a little like messenger RNA, which can code for any protein in any cell, tumor, or organ," Bathe says. "It fundamentally encodes for a protein, so it can be used across diseases, including rare diseases that may only affect a few people in the country."
Engelhardt had also worked on cssDNA as a PhD student in Munich. She met Bathe at a conference.
"We were considering collaborating on research," Engelhardt recalls. "Then Mark heard I was finishing my PhD and said, 'Wait a minute. Instead of collaborating, I should hire you.'"
Within 48 hours of submitting her PhD thesis, Engelhardt received an email asking her to apply to Bathe's lab as a postdoc. She was drawn to the position because she would be focusing on research that had the potential to help patients.
"MIT is very good at creating industry-focused postdocs," Engelhardt says. "I was inspired by the idea of doing postdoc work with the goal of spinning out a company, as opposed to doing solely academic-focused research."
Bathe and Engelhardt learned from members of the pharmaceutical industry how single-stranded DNA could help overcome limitations in gene and cell therapies. Although CRISPR-based treatments have recently been approved for a few genetic diseases, CRISPR's effectiveness has been limited by its potential toxicity and inefficient delivery to specific sites in the body. Also, those treatments can only be administered once because CRISPR often gets labeled as foreign by our immune systems and rejected from the body.
Engelhardt began exploring MIT's resources to help commercialize her research. She met Vroom through an online "founder speed dating" event at MIT. She also received support from the Venture Mentoring Service, took classes at MIT's Sloan School of Management, and worked with MIT's Industrial Liaison Program. Early on, Bathe suggested Engelhardt work with MIT's Technology License Office, something she says she tells every founder to do the moment they start thinking about commercializing their research.
In 2021, Kano won the $20,000 first place prize at the MIT Sloan Healthcare Innovation Prize (SHIP) to commercialize a new way to design and manufacture single-stranded DNA. Kano uses fermentation to produce its cssDNA less expensively than approaches based on chemical DNA synthesis.
"No one had the ability to access this type of genetic material, and so a lot of our work was around creating the highest-quality, economically scalable process to allow circular single-stranded DNA to be commercially viable," Engelhardt says.
Engelhardt and Vroom began meeting with investors as soon as Engelhardt finished her postdoc work in 2021. The founders worked to raise money over the next year while Vroom finished his MBA.
Today, Kano's circular ssDNA can be used to insert entire genes, up to 10,000 nucleotides long, into the body. Kano is planning to partner with pharmaceutical companies to make their gene therapies more targeted and potent. For instance, pharmaceutical partners could use Kano's platform to join the CD19 and CD20 genes, which are expressed in certain tumor cells, and stipulate that only if both genes bind to a cell receptor do they enter that cell's genome and make edits.
Overall, Engelhardt says working with circular single-stranded DNA makes Kano's approach more flexible than platforms like CRISPR.
"We realized working with pharmaceutical companies early on in my postdoc there was a lack of design understanding because of the lack of access to these molecules," Engelhardt says. "When it comes to gene or cell therapies, people just think of the gene itself, not the flanking sequences or anything else that goes around the gene. Now that the DNA isn't stuck in a double helix all the time, I can create small, three-dimensional structures - think loops or hairpins - that work, for example, as a binding protein that pulls it into the nucleus. That unlocks a completely new path for DNA because it makes it engineerable - not only on a structural level but also a sequence level."
Partnering for impact
To facilitate more partnerships, Kano is signing agreements with partners that give it a smaller percentage of eventual drug royalties but allow it to work with many companies at the same time. In a recent collaboration with Merck KGaA, Kano combined its circular cssDNA platform with the company's lipid nanoparticles solutions for delivering gene therapies. Kano is also in discussions with other large pharmaceutical companies to jointly bring cancer drugs into the clinic over the next two years.
"That's exciting because we'll be implementing our DNA into partners' drug system, so when they file their new drug and dose their first patients, our DNA is going to be the therapeutic information carrier for efficacy," Engelhardt says. "As a first-time founder, this is where you want to go. We talk about patient impact all the time, and this is how we're going to get it."
Kano is also developing the first databank mapping cssDNA designs to activity, to speed up the development of new treatments.
"Right now, there is no understanding of how to design DNA for these therapies," Engelhardt says. "Everyone who wants to differentiate needs to come up with a new editing tool, a new delivery tool, and there's no connecting company that can enable those areas of expertise. When partners come to us, we can say, 'The gene sequence is all yours.' But often it's not just about the sequence. It's also about the promoter or flanking sequence that allows you to insert your DNA into the genome, or that makes DNA package well into your delivery nanoparticle. At Kano, we're building the best knowledgebase to use DNA material to treat diseases."
