Only on twitter and late on Friday night and with plain English? Even the NOT reads like typical Musk tripe.

I think that’s part of it! We make cells under ludicrously hypermetabolic conditions. It’s like training at sea level and then expecting to perform at altitude!

Thanks to my incredible team and funding through the Mark Foundation, @cancerresearchinst.bsky.social, @niaidnews.bsky.social, @upmchillmancc.bsky.social and the Hillman Cancer Center. Trials of ACT with DCA conditioning are on the horizon! 13/13

Blocking the citrate carrier with benzyl-trycarboxylate completely negated the beneficial effects of DCA on T cells, suggesting a large part of metabolic reprogramming with DCA was diverting glucose-derived carbon not just to mitochondria, but also to nuclear sources for epigenetic remodeling 12/

Finally, we asked HOW mitochondrial reprogramming as altering the T cell epigenome. Carbon doesn’t always get oxidized in the mitochondria: much of it can leave as citrate through the citrate carrier (Slc25a1) where it fuels the nucleocytoplasmic pool of acetyl-CoA. 11/

Metabolomic, epigenetic, and transcriptional analyses revealed DCA induces some metabolic rewiring that allows T cells to more metabolize physiologic carbon sources present in the serum. DCA conditioning also induces changes of the epigenome, especially around stemness genes like Tcf7 and Klf2. 10/

However, cotransfer experiments of control of DCA-conditioned T cells revealed an incredible QUANTITATIVE advantage of the DCA-conditioned T cells, evident acutely after infusion (3-6 h) and did not require even antigen to be present 9/

What was really intriguing is that when we looked at the ‘per cell’ functionality of the therapeutic T cells in vivo (after infusion) there were no appreciable differences. They differentiated into effector T cells and eventually became exhausted. 8/

So, we made therapeutic murine (Pmel) or human (CD19 CAR) T cells in the presence of DCA. When used to treat tumor-bearing mice, T cells cultured in DCA were dramatically more efficacious. This occurred in the absence of lymphodepletion, in vivo vaccination, or IL-2 treatments. 7/

Targeting PDHK1 with dichloroacetate (DCA) doesn’t starve the cell of glucose, it redirects pyruvate into the mitochondria. If we activated T cells normally and expanded them in DCA, they kinda looked like in vivo T cells! High mitochondrial capacity and only minor effects on T cell expansion! 6/

Back in 2018, we identified a signaling pathway originating at the TCR that triggered aerobic glycolysis: tyrosine phosphorylation of PDHK1, an inhibitory kinase for PDH. A kinase we could inhibit! pmc.ncbi.nlm.nih.gov/articles/PMC... 5/

This is not surprising – T cell mitogens trigger aerobic glycolysis. But even 7 days after expansion, these cells were still secreting lactate like whoa. So, the big questions was: could we target aerobic glycolysis in a smart way and get in vitro T cells to behave like long-lived in vivo ones? 4/

We first compared the same T cells (OT-I) expanded in vivo (Vaccinia-OVA) to those activated and cultured in vitro. Compared metabolically, VV-OVA-primed T cells, expanding at equivalent rates, had elevated mitochondrial capacity, while in vitro expanded T cells did A LOT more glycolysis. 3/

Cellular therapies all have a common thread: they need to spend time in vitro. But we all recognize that the in vitro environment is ridiculous hypermetabolic. So with the help of Andrew Frisch and Yiyang Wang in the lab, we asked a simple question: is hypermetabolic culture also a stressor? 2/