Lasting Gammaproteobacteria profile changes characterized hematological cancer patients who developed oral mucositis following conditioning therapy

Figures & data

Table 1. Demographics of hematological cancer patients stratified per oral mucositis status post-conditioning therapy.

	T0 to TMuc^b		T0 to TYear^b
Criteria Set-All^a	Muc0-1^c	Muc2^c	Muc0-1^c	Muc2^c
Patient (M/F)	12 (8/4)	8 (3/5)	8 (5/3)	7 (4/3)
Paired sample	22	15	21	21
Age^d:
Median	56.5	56	58	48
Mean	49.8	51.1	51.9	48.3
Standard deviation	18.4	15.5	16.5	13.6
Range	25-76	23-68	25-67	23-63
Ethnicity^e:
M: C/AA	4/4	1/2	3/2	2/2
F: C/AA	4/0	3/2	3/0	1/2

^aPatient cohort (Set-All, n = 22) corresponds to hematological cancer patients undergoing conditioning therapy and having stimulated saliva samples, swabs of buccal mucosa, superficial supragingival plaque, and tongue collected for oral microbiome profiling by next-generation sequencing, pre- and post-conditioning. Demographic data are shown only for patients with successfully collected and NGS processed samples across three time points. There was similar sample type representation between Muc0-1 and Muc2 groups for the T₀ to T_Muc time period.

^bTime points were: ‘T_0ʹ (time at which cancer is present, pre-conditioning); ‘T_Muc’ (time point when patients may have developed OM WHO score 1 to 4, post-conditioning, or time point when patients did not develop OM [OM score 0] at day of transplant, day 7 and day 14 post-conditioning; ‘T_Year’ (time point one-year post-conditioning).

^cFor all three time points combined, the oral mucositis (OM) groups were: (i) patients who did not develop OM (score 0) or developed OM with score 1 (MUC_0-1), post-conditioning; (ii) patients who developed OM with score 2 (MUC₂), post-conditioning. There were 43 (= 22 + 21) and 36 (= 15 + 21) total paired sample counts T₀ to T_Muc and T₀ to T_Year combined, for MUC_0-1 and MUC₂, respectively. Overall, 114 oral samples forming pairs were collected and sequenced across the three time points. No patient developed a mucositis score of 3 or 4 for this patient cohort.

^dAverage age for T₀ to T_Muc and T₀ to T_Year patients was 50.9 and 50.2 years old, having standard deviations of 16.7 and 14.8, with ranges of 23 to 76 and 23 to 67 years of age, respectively.

^eGender and ethnicity consisted of males (M) and females (F) of Caucasian (C) or African American (AA) ethnicity.

Figure 1. Analytical design for fold changes and changes in beta-diversity of hematological cancer patients undergoing conditioning therapy with and without oral mucositis.

Table 2. Longitudinal PERMANOVA analyses of separate oral mucositis groups, Muc0-1 and Muc2, based on the PERMANOVA fixed factor ‘Time’.

Muc0-1^a	# of pts^b	Paired sample (Ct)^c	Time period^d	p-value^e
Set-All	12	22	T0-TMuc	0.003
Subset-Common	7	11	T0-TMuc	0.579
Subset-NoAntibiotics	5	11	T0-TMuc	0.094
Subset-TBI	10	6	T0-TMuc	0.355
Set-All	8	21	T0-TYear	0.291
Subset-Common	7	11	T0-TYear	0.334
Subset-NoAntibiotics	5	14	T0-TYear	0.127
Subset-TBI	6	15	T0-TYear	0.335
Muc2	# of pts	Paired sample (Ct)	Time period	p-value
Set-All	8	15	T0-TMuc	0.017
Subset-Common	6	10	T0-TMuc	0.058
Subset-NoAntibiotics	8	15	T0-TMuc	0.018
Subset-TBI	6	11	T0-TMuc	0.029
Set-All	7	21	T0-TYear	0.031
Subset-Common	6	10	T0-TYear	0.102
Subset-NoAntibiotics	6	19	T0-TYear	0.051
Subset-TBI	4	12	T0-TYear	0.156

^aPatient groups analyzed were designated as Muc0-1 and Muc2 with the first group corresponding to patients with no OM or OM score 1 and the second group representing patients with OM score 2, post-conditioning. Set-All in the Muc0-1 and Muc2 groups consisted of all patients having paired sample data (stimulated saliva, buccal mucosa, superficial supragingival plaque, or tongue) for T₀ to T_Muc [T₀-T_Muc] and T₀ to T_Year [T₀-T_Year] time periods; ‘Subset-Common’ corresponds to all patients in common (i.e., those having matched samples by sample site for all three time periods, n = 21 pairs); ‘Subset-NoAntibiotics’ represents patients who did not receive antibiotics within 2 weeks prior to sampling at T₀ pre-transplant; ‘Subset-TBI’ describes patients who received total body irradiation during conditioning.

^bNumber of patients for each patient group.

^cTotal number of paired patient samples for each group. There were 43 (= 22 + 21) and 36 (= 15 + 21) total paired sample counts, considering T₀ to T_Muc and T₀ to T_Year time periods combined, for Muc0-1 and Muc2, respectively.

^dTime points were: ‘T_0ʹ (time at which cancer is present, pre-conditioning); ‘T_Muc’ (time point when patients developed OM WHO score 1 to 2, post-conditioning, or time point when patients did not develop OM [OM score 0] at day of transplant (day 0), day 7 and day 14 post-transplant; ‘T_Year’ (time point one-year post-transplant). Analyses were performed for the time periods T₀ to T_Muc and T₀ to T_Year.

^eLongitudinal PERMANOVA analyses were performed for Muc0-1 and Muc2 groups separately, based on Bray–Curtis similarity matrices determined from square root transformed relative abundance data derived from screening of all 737 probes comprised of 620 species and 117 genus probes, using PRIMER_v7 (PRIMER-E Ltd., Ivybridge, UK). Monte-Carlo corrected p-values for the fixed factor ‘Time’ (α = 0.05) are shown. Significant p-values are highlighted (grey), marginal p-values are underlined.

Figure 2. LEfSe results of set-all Muc0-1 and Muc2 groups.

Figure 2. (Continued).

Figure 3. Average fold change (RA-FcD) difference from T₀ to T_Muc distinguishing Muc0-1 from Muc2 hematological cancer patient groups undergoing conditioning therapy.