M005670 Radiation tumor

Author

Lu Mao

Statistical analysis

Tumor growth curves were plotted by mean volume (mm\(^3\)) \(\pm\) standard deviation at each measurement occasion. Between-group comparisons were based on the average growth volume, i.e., the area under the growth curve divided by the follow-up period (Patten et al. 2022). Kaplan–Meier curves were used to estimate the survival probabilities and were compared between groups by the log-rank test. P-values \(<\) 0.05 were considered statistically significant. All analyses were performed in R version 4.3.3 (R Foundation for Statistical Computing, Vienna, Austria).

Results

Study I

T-mAb-Cetuximab (Cet)

Tumor growth and survival curves are plotted in Figure 1. Table 1 and Table 2 summarize the group-specific average tumor growth, and median survival time along with their test results.

Figure 1: Tumor growth and survival curves for Study I (cet).
Table 1: Study I (cet) 35-day average growth (mm^3) and test comparing each group to control.
Group Avg. growth (95% CI) P
No Rx 87.5 (51.6, 123.5) —
Cet+RT 34.5 (8.3, 60.7) 0.013
CP-IL2ZA+RT 26.6 (7.8, 45.5) 0.005
CP-IL2ZA+Cet 75.4 (40.3, 110.5) 0.549
CP-IL2ZA+Cet+RT -65.6 (-88.2, -43) <0.001
Additional tests on tumor growth (Group 1 vs 2).
Group 1 Group 2 Diff P
Cet+RT CP-IL2ZA+RT 7.9 (-20.5, 36.3) 0.545
Cet+RT CP-IL2ZA+Cet -40.9 (-79.3, -2.5) 0.039
Cet+RT CP-IL2ZA+Cet+RT 100.1 (70.1, 130.2) <0.001
CP-IL2ZA+RT CP-IL2ZA+Cet -48.8 (-84.8, -12.7) 0.014
CP-IL2ZA+RT CP-IL2ZA+Cet+RT 92.2 (66.6, 117.9) <0.001
CP-IL2ZA+Cet CP-IL2ZA+Cet+RT 141 (104, 178) <0.001
Table 2: Study I (cet) median survival time and log-rank test comparing each group to control.
Group Median (95% CI) P
No Rx 73 (61, —) —
Cet+RT 53 (32, —) 0.310
CP-IL2ZA+RT 61 (61, —) 0.758
CP-IL2ZA+Cet 61 (61, —) 0.031
CP-IL2ZA+Cet+RT 114.5 (113, —) 0.001
Additional tests on survival (Group 1 vs 2).
Group 1 Group 2 P
CP-IL2ZA+RT CP-IL2ZA+Cet 0.332
CP-IL2ZA+RT CP-IL2ZA+Cet+RT <0.001
CP-IL2ZA+Cet CP-IL2ZA+Cet+RT <0.001
Cet+RT CP-IL2ZA+RT 0.745
Cet+RT CP-IL2ZA+Cet 0.745
Cet+RT CP-IL2ZA+Cet+RT 0.002

Conclusion:

  • Tumor growth suppression: CP-IL2ZA+Cet+RT \(>\) CP-IL2ZA+RT \(\approx\) Cet+RT \(>\) CP-IL2ZA+Cet \(\approx\) No Rx;
  • Survival: CP-IL2ZA+Cet+RT \(>\) CP-IL2ZA+RT \(\approx\) Cet+RT \(\approx\) CP-IL2ZA+Cet \(\approx\) No Rx.

Comment: CP-IL2ZA+Cet tests significant with no Rx on survival because its survival curve is consistently lower than no Rx. But the actual difference is small.

Trastuzumab (Trast)

Tumor growth and survival curves are plotted in Figure 2. Table 3 and Table 4 summarize the group-specific average tumor growth, and median survival time along with their test results.

Figure 2: Tumor growth and survival curves for Study I (trast).
Table 3: Study I (trast) 34-day average growth (mm^3) and test comparing each group to control.
Group Avg. growth (95% CI) P
No Rx 228.4 (130.7, 326.1) —
Trast+RT 65.2 (10.7, 119.6) 0.006
CP-IL2ZA+RT 27.6 (8.7, 46.5) 0.004
CP-IL2ZA+Trast 89.7 (49.5, 129.9) 0.013
CP-IL2ZA+Trast+RT -40.5 (-69.5, -11.5) <0.001
Additional tests on tumor growth (Group 1 vs 2).
Group 1 Group 2 Diff P
Trast+RT CP-IL2ZA+RT 37.6 (-16.9, 92) 0.143
Trast+RT CP-IL2ZA+Trast -24.5 (-83.9, 34.8) 0.376
Trast+RT CP-IL2ZA+Trast+RT 105.6 (49.8, 161.4) 0.003
CP-IL2ZA+RT CP-IL2ZA+Trast -62.1 (-102.8, -21.3) 0.009
CP-IL2ZA+RT CP-IL2ZA+Trast+RT 68 (37.4, 98.7) <0.001
CP-IL2ZA+Trast CP-IL2ZA+Trast+RT 130.1 (86.6, 173.7) <0.001
Table 4: Study I (trast) median survival time and log-rank test comparing each group to control.
Group Median (95% CI) P
No Rx 66 (63, —) —
Trast+RT 68.5 (30, —) 0.420
CP-IL2ZA+RT 68 (66, —) 0.427
CP-IL2ZA+Trast 61 (61, —) 0.601
CP-IL2ZA+Trast+RT 112.5 (110, —) 0.001
Additional tests on survival (Group 1 vs 2).
Group 1 Group 2 P
Trast+RT CP-IL2ZA+RT 0.549
Trast+RT CP-IL2ZA+Trast 0.410
Trast+RT CP-IL2ZA+Trast+RT <0.001
CP-IL2ZA+RT CP-IL2ZA+Trast 0.967
CP-IL2ZA+RT CP-IL2ZA+Trast+RT <0.001
CP-IL2ZA+Trast CP-IL2ZA+Trast+RT <0.001

Conclusion:

  • Tumor growth suppression: CP-IL2ZA+trast+RT \(>\) CP-IL2ZA+RT \(>\) CP-IL2ZA+Trast \(\approx\) Trast+RT \(>\) No Rx;
  • Survival: CP-IL2ZA+Trast+RT \(>\) the rest.

Study II

Tumor growth and survival curves are plotted in Figure 3. Table 5 and Table 6 summarize the group-specific average tumor growth, and median survival time along with their test results.

Figure 3: Tumor growth and survival curves for Study II.
Table 5: Study II 35-day average growth (mm^3) and test comparing each group to control.
Group Avg. growth (95% CI) P
PBS 282 (120.7, 443.3) —
Cet+RT 109.7 (65.4, 153.9) 0.041
CP-IL2ZA+RT 79.2 (47.7, 110.6) 0.021
CP-IL2ZA+Cet 184.6 (115.6, 253.6) 0.223
3xTx 4.6 (-16, 25.1) 0.005
Additional tests on tumor growth (Group 1 vs 2).
Group 1 Group 2 Diff P
Cet+RT CP-IL2ZA+RT 30.5 (-19.8, 80.8) 0.214
Cet+RT CP-IL2ZA+Cet -74.9 (-151.8, 1.9) 0.055
Cet+RT 3xTx 105.1 (58.7, 151.5) <0.001
CP-IL2ZA+RT CP-IL2ZA+Cet -105.4 (-178.4, -32.4) 0.008
CP-IL2ZA+RT 3xTx 74.6 (38.8, 110.4) <0.001
CP-IL2ZA+Cet 3xTx 180 (109.3, 250.7) <0.001
Table 6: Study II median survival time and log-rank test comparing each group to control.
Group Median (95% CI) P
PBS 55.5 (50, —) —
Cet+RT 75 (61, —) 0.003
CP-IL2ZA+RT 75 (71, —) <0.001
CP-IL2ZA+Cet 61 (61, —) 0.016
3xTx 103 (89, —) <0.001
Additional tests on survival (Group 1 vs 2).
Group 1 Group 2 P
CP-IL2ZA+RT CP-IL2ZA+Cet 0.357
CP-IL2ZA+RT 3xTx <0.001
CP-IL2ZA+Cet 3xTx <0.001
Cet+RT CP-IL2ZA+RT 0.846
Cet+RT CP-IL2ZA+Cet 0.287
Cet+RT 3xTx 0.003

Conclusion:

  • Tumor growth suppression: 3xTx \(>\) CP-IL2ZA+RT \(\approx\) Cet+RT \(>\) CP-IL2ZA+Cet \(\geq\) PBS;
  • Survival: 3xTx \(>\) CP-IL2ZA+RT \(\approx\) Cet+RT \(>\) CP-IL2ZA+Cet \(>\) PBS.

Study III

(i)

Tumor growth and survival curves are plotted in Figure 4. Table 7 and Table 8 summarize the group-specific average tumor growth, and median survival time along with their test results.

Figure 4: Tumor growth and survival curves for Study III (i).
Table 7: Study III (i) 35-day average growth (mm^3) and test comparing each group to control.
Type Group Avg. growth (95% CI) P
ER PBS 155.2 (78.7, 231.7) —
3xTx -6.7 (-47.4, 33.9) 0.001
WT PBS 296.1 (157.8, 434.5) —
3xTx 169.2 (110.1, 228.2) 0.072
Table 8: Study III (i) median survival time and log-rank test comparing each group to control.
Group Median (95% CI) P
PBS 41 (41, —) —
3xTx 61 (53, —) <0.001

Conclusion:

  • Tumor growth suppression: 3xTx \(>\) PBS (borderline significant for WT);
  • Survival: 3xTx \(>\) PBS.

(ii)

Tumor growth and survival curves are plotted in Figure 5. Table 9 and Table 10 summarize the group-specific average tumor growth, and median survival time along with their test results.

Figure 5: Tumor growth and survival curves for Study III (ii).
Table 9: Study III (ii) 35-day average growth (mm^3) and test comparing each group to control.
Type Group Avg. growth (95% CI) P
ER PBS 246.8 (103.2, 390.4) —
CRT 135.5 (97.5, 173.6) 0.105
3xTx -7.6 (-20.7, 5.5) 0.006
3xTx+CRT -10.7 (-32.2, 10.9) 0.005
WT PBS 347.1 (269.7, 424.5) —
CRT 350.6 (284.4, 416.7) 0.930
3xTx 259.6 (190.8, 328.5) 0.060
3xTx+CRT 110.9 (80.6, 141.1) <0.001
Additional tests on tumor growth (Group 1 vs 2).
Type Group 1 Group 2 Diff P
ER CRT 3xTx 143.2 (105.9, 180.5) <0.001
CRT 3xTx+CRT 146.2 (107.7, 184.7) <0.001
3xTx 3xTx+CRT 3 (-20.7, 26.8) 0.788
WT CRT 3xTx 90.9 (8.1, 173.8) 0.034
CRT 3xTx+CRT 239.7 (174.3, 305.1) <0.001
3xTx 3xTx+CRT 148.8 (77.2, 220.3) <0.001
Table 10: Study III (ii) median survival time and log-rank test comparing each group to control.
Group Median (95% CI) P
PBS 47 (42, —) —
CRT 53 (52, —) 0.010
3xTx 60 (57, —) <0.001
3xTx+CRT 83 (74, —) <0.001
Additional tests on survival (Group 1 vs 2).
Group 1 Group 2 P
CRT 3xTx <0.001
CRT 3xTx+CRT <0.001
3xTx 3xTx+CRT <0.001

Conclusion:

  • Tumor growth suppression
    • ER: 3xTx+CRT \(\approx\) 3xTx \(>\) CRT \(>\) PBS (borderline significant)
    • WT: 3xTx+CRT \(>\) 3xTx \(>\) CRT \(\approx\) PBS
  • Survival: 3xTx+CRT \(>\) 3xTx \(>\) CRT \(>\) PBS.

Study IV

Tumor growth and survival curves are plotted in Figure 6. Table 11 and Table 12 summarize the group-specific average tumor growth, and median survival time along with their test results.

Figure 6: Tumor growth and survival curves for Study IV.
Table 11: Study IV 35-day average growth (mm^3) and test comparing each group to control.
Group Avg. growth (95% CI) P
PBS 403.5 (255.9, 551) —
3xTx-NK_dep 226.7 (142.9, 310.5) 0.028
3xTx-Mo_dep 260.8 (185.7, 335.9) 0.060
3xTx-Isotype Control 80.8 (53.9, 107.7) 0.002
Additional tests on tumor growth (Group 1 vs 2).
Group 1 Group 2 Diff P
3xTx-NK_dep 3xTx-Mo_dep -34.1 (-131.8, 63.6) 0.454
3xTx-NK_dep 3xTx-Isotype Control 145.9 (62.1, 229.7) 0.005
3xTx-Mo_dep 3xTx-Isotype Control 180 (104.8, 255.2) <0.001
Table 12: Study IV median survival time and log-rank test comparing each group to control.
Group Median (95% CI) P
PBS 46 (42, —) —
3xTx-NK_dep 56 (56, —) 0.010
3xTx-Mo_dep 56 (56, —) 0.003
3xTx-Isotype Control 72 (72, —) <0.001
Additional tests on survival (Group 1 vs 2).
Group 1 Group 2 P
3xTx-NK_dep 3xTx-Mo_dep 0.393
3xTx-NK_dep 3xTx-Isotype Control <0.001
3xTx-Mo_dep 3xTx-Isotype Control 0.001

Conclusion:

  • Tumor growth suppression: 3xTx-Isotype Control \(>\) 3xTx-Mo_dep \(\approx\) 3xTx-NK_dep \(>\) PBS;
  • Survival: 3xTx-Isotype Control \(>\) 3xTx-Mo_dep \(\approx\) 3xTx-NK_dep \(>\) PBS.

Additional mice review experiment

Depletion study

Tumor growth and survival curves are plotted in Figure 7. Table 13 summarizes the group-specific average tumor growth with test results.

Figure 7: Tumor growth and survival curves for depletion study.
Table 13: Depletion study 35-day average growth (mm^3) and test comparing each group to control.
Group Avg. growth (95% CI) P
Control/PBS 464.5 (329.2, 599.8) —
3xTx-CD115 76.6 (18.8, 134.4) <0.001
3xTx-NK 144 (54.6, 233.4) <0.001
3xTx -17.1 (-59, 24.8) <0.001
Additional tests on tumor growth (Group 1 vs 2).
Group 1 Group 2 Diff P
3xTx-CD115 3xTx-NK -67.4 (-163.9, 29.2) 0.151
3xTx-CD115 3xTx 93.7 (29.1, 158.3) 0.008
3xTx-NK 3xTx 161.1 (68.9, 253.3) 0.003

Two-tumor study

Tumor growth and survival curves are plotted in Figure 8. Table 14 summarizes the group-specific average tumor growth with test results.

Figure 8: Tumor growth and survival curves for two-tumor study.
Table 14: Two-tumor study 35-day average growth (mm^3) and test comparing each group to control.
Group Avg. growth (95% CI) P
Control/PBS 464.5 (329.2, 599.8) —
3xTx 34 (-17, 84.9) <0.001
3xTx - no RT 1.3 (-1, 3.5) <0.001

References

Patten, Luke W., Patrick Blatchford, Matthew Strand, and Alexander M. Kaizer. 2022. “Assessing the Performance of Different Outcomes for Tumor Growth Studies with Animal Models.” Animal Models and Experimental Medicine 5 (3): 248–57. https://doi.org/10.1002/ame2.12250.