ARW's CGT+RNA Manufacturing Must-Reads (Pizza Cake Edition!)

By Anna Rose Welch, Editorial & Community Director, Advancing RNA

Day-in and day-out, I write, read, listen to, and watch as much content as I can about CGT and RNA therapy manufacturing, in particular, and/or other ATMP industry-related topics that you should at least be aware of in the manufacturing facility. Every two weeks, I compile the articles and industry updates I think are most worthy of your time into an unconventional newsletter format (below) and send them out via email.

On April 20, we celebrated two of my absolute favorite things in this world: Pizza & Cake. Put the two together and you get National Pizza Cake Day — which, yes, is an actual thing. In fact, you could make one tonight for dinner if you were so inclined, seeing as your must-reads are part “instruction manual” on how to construct one of these works of art while also showcasing the equally artful technical happenings across the CGT world. Bon Appetit.

[REGULATORY]
Step 1: Preheat the oven.

• Say “duh” all you want — I see you over here with the rest of us (a.k.a. everyone) who forgets this critical step.
• Luckily, we have regulators to help us sort out which of our efforts remain only half-baked. For those of you looking to get more regular/structured feedback for your therapies, EMA announced a few valuable additions to its Priority Medicines (PRIME) Scheme, which Daniela Drago clearly outlines in her recent LinkedIn post.
• For those of us trying to figure out how exactly to even turn on our ovens/start developing an ATMP, CardinalHealth has put together a list of six tips to help take advantage of and get the answers/help you crave from the regulators during an INTERACT meeting.

[PROCESS DEVELOPMENT]
Step 2-3: Gather your highest-quality ingredients and cooking supplies. Then throw everyone else out of the house.

• You’re not just making a basic pizza here, you’re making a pizza cake. Ramp up the class & focus a little bit, please.
• The process with which you assemble a pizza cake is just as multifaceted as establishing a commercial-ready mRNA drug product manufacturing platform. I can’t say I got any “secret [pizza] sauce” recipes out of Nutcracker Therapeutics’ COO John Stubenrauch or Tessera Therapeutics’ COO Hari Pujar. But I daresay we all enjoyed digging into the operational best practices to employ in each layer of the mRNA manufacturing practice in this latest BioProcess Online Live event (i.e., transition from R&D to CMC; IVT optimization; downstream processing).

[ANALYTICS]
Step 4: Identify toppings and ensure your supply will be sufficient for this “masterpieczza.”

• If you’re (rightfully) putting pineapple on and in this pizza, there better be no such thing as a layer with only half, partial, or NO pineapple on it. Consistency is key.
• Speaking of leaving no layer un-pineappled: full/empty AND partial AAV capsids have been getting quite a bit of chatter the last few months. In particular, how we characterize partial capsids (and the other species residing within them) is growing increasingly important and more regularly studied. Two recent publications include:
  • European Biophysics Journal: AAV Analysis By Sedimentation Velocity Analytical Ultracentrifugation: Beyond Empty and Full Capsids
  • Analytical Chemistry: Analysis of AAV-Extracted DNA by Charge Detection Mass Spectrometry Reveals Genome Truncations (Not open access)
    • Full-text available via this LinkedIn post

[MANUFACTURING REPRODUCIBILITY/INFRASTRUCTURE]
Steps 5-10: Make one pizza. Then stack five more on top of that.

• Because one layer is never enough whether we’re talking pizza or manufacturing capabilities.
• Though Moderna wasn’t constructing the “leaning tower of pizza,” I loved hearing more about the company’s efforts during the pandemic to scale-out their process to meet global need. In part 2 of my three-part series, I explore how Moderna’s journey exemplified our tried-and-true motto, “begin with the end in mind.”
• I’ve made the argument before that the CGT manufacturing space, like a rainforest (or a good friend who shares their pizza cake), has become infinitely more “symbiotic.”
• That’s why I was particularly intrigued by this brief article about Myeloid Therapeutics’ new mRNA manufacturing facility. Specifically, the breakdown of how that capacity is going to be used suggests a seemingly similar symbiotic future taking shape in the burgeoning mRNA manufacturing space.

[PHYSICIAN/PATIENT CENTRICITY]
Step 11: Dish it all out.

• Reminder: To avoid an abysmal & unconscionable eating experience, please bake the pizza cake before consuming.
• If you think the terms “abysmal & unconscionable” are overly dramatic in the context of pizza, please be aware that these words are a direct quote from a physician describing the autologous cell therapy manufacturing process in this recent article.
  • Overall, as we strive to move these therapies into solid tumors and turn them into second-/first-line therapies, the article above is a goldmine of how physicians are considering CAR-Ts against competitor therapies (i.e., bispecific antibodies) — and in some instances, they’re not mincing words.
  • Not only do we need to figure out solutions to the manufacturing shortcomings of autologous therapies, but I think this re-emphasizes a particularly salient point from a conference earlier this year (i.e., “Hot Take” #2) that enabling physicians to engage with your product prior to the bedside is a foundational part of scaling your cell therapy operations.
  • If you’re trying to imagine what the future of a more efficient and patient- & physician-centric CT industry might look like, Friends of Cancer Research will be hosting a hybrid event on May 22, 2023 featuring an impressive line-up of industry, government, FDA, and physician speakers on the future development, clinical, & policy landscapes for CTs. You can register for free here.

[NON-VIRAL DELIVERY]
Step 12: Dig in — preferably with a fork & knife.

• Or if you must, eat with your hands — but first, please wash them to ensure this epicurean adventure remains a “non-viral” delivery from plate to mouth.
• There have been a lot of LNP and polymer nanoparticle (PNP) cooks in the kitchen over the past two years. Bless the good folks at Aust Business Solutions for serving up this open access Excel list comprising 180+ publications in the LNP and PNP fields from 2020-2022.

[CRYOPRESERVATION/mRNA STABILITY]
Step 13: Put all the leftovers in the fridge.

• Or maybe breakfast = pizza cake left on the counter overnight. (I’m judging you, though.)
• The beauties of a future with mRNA that doesn’t require refrigeration/freezing have been explored in a handful of recent publications on mRNA-LNP stability. We can slice this massive topic into a couple of different pieces, starting first with understanding the important role lipid & non-lipid excipients play in a product’s deliverability and stability.
• It’s perhaps incorrect to say freeze drying is a “hot” topic in the mRNA-LNP formulation world, but this recent publication on continuous freeze-drying of mRNA LNPs definitely heats things up. (Full text available)
• This publication is a formidable buffet of all the factors that impact mRNA vaccine stability (i.e., mRNA structure, excipients, LNPs, manufacturing processes, temperature/physical stress). However, it also gives us a taste of current global regulatory guidance on mRNA vaccine stability studies.