BioProject

Patients with multiple myeloma (MM) routinely receive mRNA-based vaccines to reduce COVID-19-related mortality, but whether disease- and therapy-related alterations in immune cells and cytokine-responsiveness contribute to the observed heterogeneous vaccination responses is unclear. Thus, we analyzed PBMCs from patients with MM during/after SARS-CoV-2 vaccination and breakthrough infection (BTI) using combined whole-transcriptome and surface proteome single-cell profiling with functional serological and T-cell validation in additional 58 patients with MM. Our results demonstrate that vaccination responders showed a significant overrepresentation of cytotoxic CD4+ T-cells and mature CD38+ NK-cells expressing FAS+/TIM3+ with a strong enrichment for cytokine-responsiveness such as type-I-interferon-, IL-12- and TNF-alpha-mediated signaling. Patients with MM experiencing BTI developed higher serological and cellular responses and displayed similar cytokine-responsive immune cell patterns as observed in vaccination responders. Finally, these results expand our understanding of molecular and cellular patterns associated with immunization responses and may benefit the design of improved vaccination strategies in immunocompromised patients. Overall design: For single-cell CITE sequencing of T-, B-, NK- and NKT-cells of patients with Multiple Myeloma (MM) and age-matched healthy controls, we selected 11 representative individuals from our previously published cohort (Enssle JC, Cancer Cell, 2022) resulting in 24 samples. Samples were obtained from serological and T-cell responders and non-responders after the 2nd (TP3) and 3rd (TP5) vaccination. To limit potential disease- or treatment-related bias, we selected patients which are representative of the group that was in the 1st line of treatment for MM, in at least very-good-partial-remission (VGPR) and without corticosteroid treatment 14 days before and after vaccination. We also included samples from 2 healthy controls (HC) after their respective 2nd and 3rd vaccination. As some patients with MM from our main cohort exhibited a breakthrough infection (BTI) after the 3rd vaccination at timepoint of data cut-off, we additionally included 2 full responders and 2 full non-responders MM patients together with 2 HC which all developed a BTI after the 3rd vaccination. Here, we subjected samples after the 3rd vaccination and after BTI (TP6) for CITEseq analysis. One patient with MM donated samples after TP3, TP5 and BTI. >>>Raw data for human samples not available due to patient privacy concerns<<<