1. *
OS >3yr with
No Next Line * * * * *
Supplemental Figure 1. Schematic presentation of the analyzed parameters in the correlative cohort per progression survival (PFS). TMB: total mutational burden. PR: partial response. SD: stable disease. PD: progressive disease. △: mutation.

2. Supplemental Figure 2. Progression Free Survival (PFS) and subgroup survival analysis of the correlative cohort (N=38) by Log-rank (Mantel-Cox) test.
PFS By PD-L1 categories ( 0-49% n=23 vs % n=9, p<0.001)
PFS By TMB percentile (<25th percentile n=6 vs 25-50th percentile n=7 vs 50-75th percentile n=6 vs >75th percentile n=6, p=0.142)
PFS By CD8 categories (0-5% n=18 vs 6-25% n=14, p=0.037)
PFS By CD4 categories (0-5% n=21 vs 6-25% n=9 vs >25% n=2, p=0.123)
P<0.001
P=0.142
P=0.037
P=0.123

3. Supplemental Figure 3. Association among the four correlative parameters.

4. Supplemental Table 1. Association of the four correlative parameters and with other clinical characteristics.
TMB
PD-L1
CD8
CD4
Smoking
EGFR
Squamous
Spearman
1.000
0.191
0.125
0.072
.517**
-0.333
-0.221
p-value
0.406
0.591
0.758
0.008
0.104
0.289 N 25 21
.660**
.487**
.366*
-0.201
.395*
<0.001
0.005
0.040
0.271
0.025 32
.396*
0.342
-0.287
0.121
0.055
0.111
0.508
-0.003
0.177
-0.031
0.985
0.334
0.865

5. Supplemental Table 2. Correlation of patient characteristics and ORR per irRC (PR vs SD/PD).
ORR (irRC)
Median (range)
PR (n=16)
SD/PD (n=22)
p-value
Test
# missing
TMB
189 ( )
55 (48-173)
0.077
Wilcx 13
PD-L1
66 (29-81)
15.3 (4-32)
0.002 6
CD8
8 (4-11)
3 (1-7)
0.017
CD4
8 (1.5-12)
2.3 (1-5)
0.168
Age
68 (61-75)
66 (59-74)
0.455
Male
9 (56.3%)
13 (59.1%) 1
Fish.
EGFR Mutations
3 (21.4%)
6 (27.3%)
ALK
0 (0.0%)
1 (4.5%) 5
EGFR or ALK
7 (31.8%)
0.706 2
Ever smoker
12 (54.5%)
Squamous
3 (18.8%)
5 (22.7%)
>=One Line Therapy
11 (68.8%)
19 (86.4%)
0.181

6. Supplemental Table 3. Patient characteristics (as categorical variables) in the correlative cohort per prior lines of therapy.
No prev. line (n=8)
>= 1 line (n=30)
p-value
TMB
0.785
<25 percentile
1 (14.3%)
5 (27.8%)
25-50
2 (28.6%)
50-75
3 (42.9%)
3 (16.7%)
>75 percentile
PD-L1
0.238
<1
1 (16.7%)
0 (0.0%)
1-49
4 (66.7%)
18 (69.2%)
>49
1 (6.7%)
8 (30.8%)
CD8
0.030
0-5%
17 (65.4%)
5-25%
5 (83.3%)
9 (34.6%)
CD4
0.305
16 (61.5%)
>25%
2 (7.7%)

7. Supplemental Table 4. PFS and OS of correlative characteristics (as categorical variables) by year.
Median (range)
All correlative cohort (N=38)
PFS <= 1Yr (n=27)
PFS 1yr-2yr (n=5)
PFS 2-3yr (n=4)
PFS >3yr (n=2)
p-value
TMB
0.021
<25 percentile
6 (15.8%)
5 (31.3%)
1 (25.0%)
0 (0.0%)
25-50
7 (18.4%)
1 (50.0%)
50-75
>75 percentile
1 (6.3%)
3 (100.0%)
PD-L1
0.002
<1
1 (2.6%)
1 (4.0%)
1-49
22 (57.9%)
20 (80.0%)
2 (66.7%)
>49
9 (23.7%)
4 (16.0%)
1 (33.3%)
1 (100.0%)
CD8
0.093
0-5%
18 (47.4%)
16 (64.0%)
5-25%
14 (36.8%)
9 (36.0%)
CD4
0.095
21 (55.3%)
18 (72.0%)
6 (24.0%)
>25%
2 (5.3%)
Median (range)
All correlative cohort (N=38)
OS <= 1Yr (n=16)
OS 1yr-2yr (n=9)
OS 2-3yr (n=6)
OS >3yr (n=7)
p-value
TMB
0.662
<25 percentile
6 (15.8%)
1 (11.1%)
2 (40.0%)
1 (20.0%)
2 (33.3%)
25-50
7 (18.4%)
5 (55.6%)
0 (0.0%)
1 (16.7%)
50-75
2 (22.2%)
>75 percentile
PD-L1
0.003
<1
1 (2.6%)
1 (6.3%)
1-49
22 (57.9%)
14 (87.5%)
4 (66.7%)
1 (25.0%)
3 (50.0%)
>49
9 (23.7%)
1 (33.3%)
3 (75.0%)
CD8
0.048
0-5%
18 (47.4%)
12 (75.0%)
2 (50.0%)
5-25%
14 (36.8%)
4 (25.0%)
CD4
0.333
21 (55.3%)
4 (35.0%)
>25%
2 (5.3%)

8. Supplemental Table 5. Characteristics as continuous variables of long term benefiters (Overall survival > 3 years with no intervening therapies) vs all other patients in the correlative cohort.
Clinical characteristics Median (range)
LTB (n=5)
All others (n=33)
p-value
TMB
173 ( )
83 ( )
0.530
CD8
9.5 (5.5-15)
4 (2-8.5)
0.106
PD-L1
72 (55-77)
16 ( )
0.029
CD4
2 ( )
3.2 (1-8)
0.763
Age
59 (59-68)
68 (60-74)
0.424
Male
4 (80.0%)
18 (54.5%)
0.374
EGFR Mutations
0 (0.0%)
9 (28.1%)
0.553
ALK Mutation/Translocation
1 (3.4%)
1.000
EGFR or ALK Mutation
10 (31.3%)
0.559
Ever smoker
3 (60.0%)
Squamous
1 (20.0%)
7 (21.2%)
At Least One Line Therapy
27 (81.8%)
0.279