Microglia normally gobble up and break down Aβ. However, in Alzheimer’s disease, an altered inflammatory state causes them to stop clearing the aggregated peptide. How does this happen, and can it be stopped? Junying Yuan of Harvard Medical School, Boston, blames the microglial enzyme RIPK1, and believes that blocking it may help return microglia to their normal state. According to a paper Yuan and colleagues published September 13 in the Proceedings of the National Academy of Sciences, the kinase appears to set off transcriptional changes that cripple the microglial lysosome system. The cells start producing new gene products, some characteristic of the recently identified disease-associated microglia (DAM) surrounding plaques in AD model mice (Keren-Shaul et al., 2017; Tay et al., 2017). Genetically deleting or pharmacologically inhibiting RIPK1 both sped up Aβ clearance and improved memory in an AD mouse model. The findings lay the groundwork for a new treatment for AD, and, since RIPK1 has been implicated in amyotrophic lateral sclerosis (ALS) and multiple sclerosis (MS), for those diseases as well, the authors believe.
“It’s the first paper that shows blocking RIPK1 alleviates the inflammatory response, reduces plaque, and improves behavior in AD mice,” Yuan said. “It points out directly the beneficial effects of inhibiting RIPK1 for the treatment of multiple diseases characterized by inflammation and cell death.”
“A crucial role of RIPK1 in the switch between phagocytosis and inflammatory capacity in microglia is novel and exciting,” said Marco Colonna, Washington University in St. Louis. “The authors convincingly show that inhibition of RIPK1 increases the turnover of Aβ1–42 and reduces production of proinflammatory cytokines by microglia.”
“Microglial lysosome biology is poised to become the next hot topic in Alzheimer’s research,” said Terrence Town, University of Southern California, Los Angeles, who was not involved in the study. “A lot of recent data are pointing to failure of the lysosome in microglia and other innate immune cells as the problem in AD, and rebalancing that as the way forward.”
RIPK1, short for Receptor-interacting protein 1 kinase, gets induced in response to the inflammatory signals tumor necrosis factor (TNFa) and ligands of the toll-like receptor (TLR) family. It causes an inflammatory response, controls inflammation-induced cell death (necroptosis), and leads to some forms of apoptosis (for a review, see Ofengeim and Yuan, 2013). The kinase has already been shown to mediate the necrotic cell death and inflammation observed in amyotrophic lateral sclerosis and multiple sclerosis (see February 2017, September 2016 news; Ito et al., 2016; Ofengeim et al., 2015). Since inflammation also plays a big role in Alzheimer’s disease, Yuan wondered if RIPK1 was involved there, as well. They also wondered how it might affect disease pathology.
To find out, co-first authors Dimitry Ofengeim and Sonia Mazzitelli first peered into postmortem human brains and found more phosphorylated RIPK1 in slices from AD patients than controls. This implied that the kinase was activated in the disease. That RIPK1 co-localized with microglial markers suggested that it was expressed primarily in these cells. The same proved true in the brains of six-month-old APP/PS1 mice, where RIPK1 appeared in microglia, particularly those surrounding plaques, but not in astrocytes or neurons (see image above).
What did the kinase do? The authors tested this in APP/PS1 mice by adding to their drinking water a RIPK1 inhibitor the group had previously developed called necrostatin-1 (Nec-1s). After a month, the treated mice had fewer plaques and less soluble and insoluble Aβ in the brain. What’s more, whereas five-month-old APP/PS1 mice scurried around an open field in a hyperactive state, a month of Nec-1s treatment calmed them down. The researchers also examined spatial memory with a T-shaped water maze, where mice are trained to find a hidden platform at the end of one arm, then retrained to find it in another. At five months, APP/PS1 mice had trouble learning a new platform location, but a month of Nec-1s restored their performances to match those of wild-type mice.
The kinase activity of RIPK1 likely lay at the root of these effects, as APP/PS1 mice that expressed the protein with a mutated, inactive kinase region reaped the same physiological and behavioral benefits of receiving Nec-1s. Together the data suggested that thwarting RIPK1 action lightened amyloid pathology and subsequent symptoms.
How does a microglial kinase do this? The researchers added Aβ1-42 to microglia isolated from wild-type and kinase-dead knock-in mice. Wild-type cells bumped up production of the inflammatory cytokines TNFa and IL6, mutant cells less so. Wild-type microglia pretreated with Nec-1s also produced less TNFα and IL6.
Intriguingly, microglia lacking RIPK1 action better digested synthetic Aβ1-42 oligomers. What whetted their appetites? Analyzing the microglial transcriptomes, Ofengeim and Mazzitelli found that one of the proteins upregulated by RIPK1 was cystatin F. Encoded by the Cst7 gene, cystatin F inhibits the endosomal/lysosomal system. Microglia that expressed Cst7 surrounded plaques in APP/PS1 mouse and AD postmortem tissue.
RIPK1-regulated genes were also altered in microglia from the 5XFAD model of AD, the SOD1G93A model of ALS, and aging microglia, implying that RIPK1 went into overdrive there, as well. One of those genes was CH25h, an enzyme on the cell surface responsible for cholesterol and lipid metabolism. CH25h is an AD GWAS hit (Wollmer, 2010).
Cst7 recently drew notice as a marker of DAM cells that surround plaques in 5xFAD AD mouse models. The results suggest that active RIPK1 triggers microglia to adopt a DAM state, which slows the ability to break down Aβ and raises interest in RIPK1 inhibition as a new treatment approach for AD, the authors write.
Researchers at Denali Therapeutics in South San Francisco picked up this idea and are testing a RIPK1 inhibitor in a Phase 1 trial of healthy volunteers in Europe. Yuan actively collaborates with the group. If the compound proves safe, trials in ALS and AD patients are next.
The current paper provides “indirect evidence that releasing the brakes on cerebral innate immunity is a valid therapeutic approach,” Town told Alzforum. Once controversial, multiple lines of evidence are now converging on this notion, he said. “What used to be a fringe theory is coming into the mainstream in a big way.” He cautioned that before concluding that the lysosomal effect occurs through Cst7, the authors need to show that blocking Cst7 has the same effect as the kinase mutant.
Salvatore Oddo, Arizona State University, Tempe, said the results jibe with those his group reported earlier this year (Caccamo et al., 2017). He suggested an acceleration of RIPK1-induced necroptosis in AD. While the current paper focuses instead on resulting inflammation, both studies converge on the idea that RIPK1 could be a therapeutic target for AD, he said. “It is likely that in AD, RIPK1 influences multiple pathways, having to do with both cell death and non-cell death mechanisms.”
Caccamo A, Branca C, Piras IS, Ferreira E, Huentelman MJ, Liang WS, Readhead B, Dudley JT, Spangenberg EE, Green KN, Belfiore R, Winslow W, Oddo S. Necroptosis activation in Alzheimer’s disease. Nat Neurosci. 2017 Sep;20(9):1236-1246. Epub 2017 Jul 24 PubMed.
Ofengeim D, Yuan J. Regulation of RIP1 kinase signalling at the crossroads of inflammation and cell death. Nat Rev Mol Cell Biol. 2013 Nov;14(11):727-36. Epub 2013 Oct 16 PubMed.
Ito Y, Ofengeim D, Najafov A, Das S, Saberi S, Li Y, Hitomi J, Zhu H, Chen H, Mayo L, Geng J, Amin P, DeWitt JP, Mookhtiar AK, Florez M, Ouchida AT, Fan JB, Pasparakis M, Kelliher MA, Ravits J, Yuan J. RIPK1 mediates axonal degeneration by promoting inflammation and necroptosis in ALS. Science. 2016 Aug 5;353(6299):603-8. PubMed.
Ofengeim D, Ito Y, Najafov A, Zhang Y, Shan B, DeWitt JP, Ye J, Zhang X, Chang A, Vakifahmetoglu-Norberg H, Geng J, Py B, Zhou W, Amin P, Berlink Lima J, Qi C, Yu Q, Trapp B, Yuan J. Activation of necroptosis in multiple sclerosis. Cell Rep. 2015 Mar 24;10(11):1836-49. PubMed.
Wollmer MA. Cholesterol-related genes in Alzheimer’s disease. Biochim Biophys Acta. 2010 Aug;1801(8):762-73. PubMed.
To view commentaries, primary articles and linked stories, go to the original posting on Alzforum.org here.
Copyright 1996–2017 Biomedical Research Forum, LLC. All Rights Reserved.