Nature | Vol 577 | 23 January 2020 | 565effect of the TLS signature using previously published^27 data from an
additional cohort of 40 patients with melanoma who were receiving
CTLA4 blockade (Fig. 3f). Previous studies have demonstrated tumour
mutational burden as a predictive biomarker for response to ICB^28.
However, in this cohort of patients treated with anti-CTLA4, the TLS
signature is independent of mutational load (Extended Data Fig. 5b).
Moreover, the TLS signature was significantly associated with overall
survival in a previously published^22 dataset of pretreatment samples
from 69 patients who were undergoing anti-PD1 monotherapy or anti-
CTLA4 and anti-PD1 combination therapy (Fig. 3g). We also observed
the predictive effect of the TLS signature in a previously published^28
dataset of pretreatment samples from 41 patients who were treated with
anti-PD1 (of whom 50% had been exposed to anti-CTLA4 before anti-PD1
treatment) (Fig. 3h, Extended Data Fig. 5c). Finally, we performed meta
Cox regression analysis across the four cohorts treated with ICB, using
multiple immune signatures: of these, our TLS signature performed
best (Extended Data Fig. 5D). The TLS signature was also independent
of tumour mutational load in the cohort treated with anti-PD1 (Fig. 3i),
consistent with previous studies that have shown that immune gene
signatures are not correlated with mutational load^29. Although we did
not observe significant differences in the TLS gene-expression score
retrieved from pretreatment biospies with regards to ‘response evalu-
ation criteria in solid tumours’ (RECIST), we observed a notable differ-
ence in RNA-seq data from on-treatment biopsies that were collected on
cycle 1 at day 29, which was confirmed in previously published cases of
patients treated with anti-PD1^30 (Extended Data Fig. 5e, f ). This indicates
that TLS functionality is induced by ICB treatment in patients with a
clinical response. To further determine the biological relevance of our
TLS signature, we applied it to RNA-seq data from 13 additional samples
of melanoma that were obtained from patients who were receiving ICB,
and performed concurrent immunostaining of CD20 and CD3. The
samples with the highest TLS gene score contained TLSs (as detected
by CD20 immunostaining), which confirms the ability of our gene sig-
nature to predict samples with TLSs (Extended Data Fig. 5g).
In conclusion, our data provide evidence that TLSs may have a key
role in sustaining an immune-responsive microenvironment. This find-
ing opens avenues for therapeutic strategies that aim at enhancing
TLS formation and function, which could result in improved clinical
outcomes and responses to cancer immunotherapy.
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availability are available at https://doi.org/10.1038/s41586-019-1914-8.
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