Posted on Authorea 18 May 2020 — CC-BY 4.0 — https://doi.org/10.22541/au.158981322.20981629 — This a preprint and has not been peer reviewed. Data may be preliminary. Hoang Hall David Zvi Yoav Osteosarcoma Cell in Expressed Receptors Variably Surface of Utility Therapeutic and Prognostic ie vlae,adfrhrivsiae saptnilnvlteaetctre sn radioimmunotherapy using cell target all therapeutic across novel overexpressed potential consistently was a (IGF2R) as 2 investigated receptor further standard factor and and patient-derived growth evaluated, using Insulin-like studied lines previously [8]. prognostication was lines OS for cell in biomarkers pattern OS expression OS receptors identifying surface cell in The have interest 7] ongoing efforts need [6, an the cooperative targets. Intergroup remains underscoring Multiple therapeutic Osteosarcoma futile, There and has is [2-5]. European approaches. alone survival the decades novel chemotherapy most 5-year by four conventional for overall fifth studies intensifying nearly The that and in the demonstrated improved trial and 2]. repeatedly EURAMOS-1 not malignancy [1, recent has adults the and bone young including 70% primary and roughly non-hematologic adolescents at common among plateaued malignancy most primary the common is (OS) Osteosarcoma survival. overall targets or attractive EFS them Introduction makes predict expression to EFS consistent samples failed of their OS also predictors value, patient-derived significant prognostic line of approaches. provide be majority cell therapeutic not to the receptors per future across did found 6 for receptors expression expression were variable All receptor positive receptors, demonstrated While 6 of Results: receptors overall the number surface tested. of cell and test. total six None event log-rank sum The EFS the lines. The Conclusion: for using cell hazard survival. of assessed overall were majority in the or expression Differences with in receptor patients expressed method. for positive variably survival for Kaplan-Meier with overall were (geoMFI) patients and the intensity (EFS) by for survival fluorescent event event-free estimated mean survival The were geometric PDGFR- control. expression The Her-2, negative receptor a to therapeutic cytometry. to positive antibodies and flow relative with calculated prognostic using labeled was offer characterized were receptor was receptors each 52) current Expression these = The whether (n VEGFR-3. evaluate samples lines. and and OS cell Patient-derived patterns (OS) expression Methods: osteosarcoma previous varying value. validate across to expression sought variable study demonstrated receptor previously factor growth (VEGFR)-3, platelet-derived receptor (Her)-2, receptor factor growth epidermal human (PDGFR)- receptors, surface cell Six Background: Abstract 2020 18, May 6 5 4 3 2 1 hlrnsOclg Group Oncology Children’s Center Group Hospital Cancer Oncology Cancer Anderson Children’s Children’s MD Center Texas Cancer of Anderson University MD Medicine Texas of of University College Einstein Albert University Yeshiva Center Medical Montefiore 1 n ai Geller David and , β, 1 nui-iegot atr1rcpo IF1) nui eetr(R,cMt n aclredteilgot factor growth endothelial vascular and c-Met, (IR), receptor insulin (IGF-1R), receptor 1 factor growth insulin-like lfUgur Elif , 5 ae Tingling Janet , 2 ra Batko Brian , 1 1 oadBarkauskas Donald , 1 oahnGill Jonathan , 1 6 igagZhang Jinghang , 3 ihe Roth Michael , 3 2 ihr Gorlick Richard , u Yang Rui , β, G-R R c-Met, IR, IGF-1R, 1 Bang , 4 , Posted on Authorea 18 May 2020 — CC-BY 4.0 — https://doi.org/10.22541/au.158981322.20981629 — This a preprint and has not been peer reviewed. Data may be preliminary. h eMIfrec eetrwsrcre n xrse eaiet h epciengtv oto safold a as control negative respective the to geometric relative (geoMFI expressed as The and defined recorded were and error. was lines change technical receptor cell each a all for to across geoMFI The receptor due each were for results were geoMFI samples the control triplicate negative that line, calculated. and likelihood cell (geoMFI) each the intensity For fluorescent diminish felt characterization. mean Molt4 to was receptor control line Her-2 order negative C243 for Additional in validation the analyzed auto-fluorescence. as however, any used tubes; for was (FMO) accounting line and control, One cell negative Minus c-Met-APC, statistically better Fluorescent a and IR-FITC, of provide receptors, use IGF-1R-FITC, to the four for precluded these control samples control. for Limited negative negative expression robust the meaningful control. a as no be negative yielded used to a deemed C243 was to line relative line Cell expressed cell VEGFR3-APC. and C243 PDGFR- lines, and the cell Her-2-PE while assayed for used all controls across negative The plotted using was analyzed was expression Data Receptor analysis. for scatter required side was Characterization and Receptor cells forward live standard single Biosciences). (BD 1,000 cells, cytometer software SSC-A/SSC- of live flow FSC-A/FSC-H, FlowJo minimum FSC-A/SSC-A, single top A follows: bench for as SSC-A/DAPI. employed digital gate were H, staining, LSRII To DAPI Dickinson CA). with protocols Becton or View, gating a (FITC) Mountain using Dickinson, fluorescein-isothiocyanate antibodies. performed (Becton isotype-matched (PE), was using analysis phycoerythrin prepared cytometry – were Flow tubes fluorophores Control three 1). (Table of (APC) one allophycocyanin close VEGFR-3, to As IR, IGF-1R, conjugated sample. Her-2, was to per antibodies PDGFR- cells anti-human cytometry. available live and flow commercially of c-MET, using using number assessed done the was and staining determine stained Cell were to MEM- cells counted with live were re-suspension million cells After 1 serum, medium. to bovine freezing fetal their remove 10% to and centrifuged and thawed were survival Cells The observations. independent Analysis as Cytometry observations. available patients treated Flow had 49 samples 50 were patients Forty-eight these 81 that patients, using analysis. leaving observations 99 data performed culture, 2 these for was in had cells Of analysis grow patient of number not One were analysis. sufficient did sample data. a of samples survival yielded each time Eighteen samples for Fifty-two the died. outcomes at analysis. had for Clinical All associated patients written only 49 approval. informed biorepository. and and following (IRB) survived, investigators (COG) banked Board the Group were Review to and Oncology blinded Institutional OS Children’s localized and the high-grade consent from with patient patients obtained from were originated lines samples cells OS Ninety-nine expression lines receptor Cell surface receptor of of pattern pattern Methods variable the & the Materials Does Does (1) (2) questions: following utility? findings? the prognostic reported provide address to previously of to as subsets was corroborate given study for expression this utility was of therapeutic Consideration purpose or The prognostic expressed. either variably yield be could platelet- tumors. receptors to OS (HER)-2, these found receptor across were factor expression growth (VEGFR)-3 whether epidermal receptor Human factor including (PDGFR)- growth receptors receptor of factor group growth second derived A 10]. [9, β eetr nacrac ihmnfcueisrcin Tbe1.Ec antibody Each 1). (Table instructions manufacture with accordance in receptors positive /geoMFI β, G-R nui eetr(R,cMt n aclrendothelial vascular and c-Met, (IR), receptor insulin IGF-1R, negative .Arcpo a osdrdpstv ftefl change fold the if positive considered was receptor A ). 2 - β Ewr hi epcieioyemthdcontrols, isotype-matched respective their were PE α media Posted on Authorea 18 May 2020 — CC-BY 4.0 — https://doi.org/10.22541/au.158981322.20981629 — This a preprint and has not been peer reviewed. Data may be preliminary. nlzdimnhsohmclepeso fHr2i 1tmrsmlsfo ainswt non-metastatic al. with et Akatsuka patients controversial. from remains samples and tumor past 81 the in in deviation debated Her-2 been of standard has expression OS immunohistochemical and in analyzed Her-2 for of mean significance value account statistical prognostic achieve a to The survival. to variation Table overall failed (Supplementary reported of expression or studies findings, EFS coefficient both reported predicting the in initially in receptors these calculating al. all corroborating when across Despite However exists et 1). clearly study. expression Hassan variable current differences, the these in reported [8]. those results PDGFR- reported 7). geoMFI & Her-2, 6 previously (Tables survival study firming overall or this EFS of receptors In positive predictors lines of significant cell number be total 3 to survival. and respective them only receptors the overall Discussion find 6 to while with not relation the receptors, associated did with of events each significantly line 6 survival for cell overall be of analyses and per to ratio 2 EFS Hazard found with for patients was expressed. positive those receptors 0.0192) summarize be 5 = and to (p 3). 4 noted Tables Table diagnosis 3, were at (Figures (Figure status lines variability receptors less Metastatic cell 6 demonstrate all of which for majority c-MET, expression and The positive very VEGFR-3, or demonstrated PDGFR- no IR, 2). exhibited and IGF-1R, & which PDGFR- Her-2 to C331, 76.5%, 1 negative compared and in receptors. C232 their lines, IGF-1R lines to 6 cell cell 76.5%, relative all all included interest in Outliers for of c-MET 62.7%. receptors expression lines, in the low cell VEGFR-3 express of and to in 66.7%, 98% found in summarized in IR were are expressed lines receptors (geoMFI was cell respective change Her-2 of the control; fold majority and the The line 1, shown. cell Figure is each In for PROC geoMFI 2. using control Table positive SAS9.4 negative of and in number geoMFI done The total were the Analyses as well test. as log-rank cell Results survival receptors, the per overall 6 receptors using and FREQ. the positive event assessed PROC of of EFS were and each number for line, LIFETEST hazard total in cell in the patients per Differences as negative receptors well and method. as Meier positive Kaplan receptors, for at the 6 event alive by the were estimated for they were survival if line, overall contact and last either EFS at until The enrollment censored last from considered at days were as censored patients defined considered an was time. Osteosarcoma were survival either groups patients that Overall contact. to EFS, the event. enrollment last For an among from or contact. experience compared days death not last as were did to defined survival they or was if overall death) contact survival and or free (EFS) progression, Event survival (relapse, variables. event free demographic event the using variable of by continuous defined terms a as in also outcome and The enrollment years) at [?]10 Age (0-9, variable proportions. was categorical checked of a was data test as characteristics conditional follow-up the manner of exact of this pair the source in each by checked and the population was analytic population was the analytic that within the study association in for COG inclusion the with association plus for interest the checked line. representing of cell variables characteristic given binary demographic a were Each for six receptors first positive The of PDGFR- number analyzed. Her-2, total were of: variables absence seven or line, presence cell absence. COG or each presence For their indicating line, cell Analysis every Statistical in VEGFR3 and cMet, 50 IR, the IGF-1R, above was geoMFI in t ts raoewyaayi fvrac AOA,a appropriate. as (ANOVA), variance of analysis one-way a or -test positive /geoMFI negative th o h i eetr cosalcl ie uhdffrn naslt au than value absolute in different much lines cell all across receptors six the for ecnieo ellns iayvralswr sindfrHr2 PDGFR- Her-2, for assigned were variables Binary lines. cell of percentile β, G-R R -e,adVGR3wr aibyepesd reaf- expressed, variably were VEGFR-3 and c-Met, IR, IGF-1R, β, G-R R -e,adVGR3 h eet aibewsthe was variable seventh The VEGFR-3. and c-Met, IR, IGF-1R, positive 3 /geoMFI β xrsindmntae oevraiiyacross variability more demonstrated expression negative o ahrcpo cosalcl lines cell all across receptor each for ) β n70.5%, in β, Posted on Authorea 18 May 2020 — CC-BY 4.0 — https://doi.org/10.22541/au.158981322.20981629 — This a preprint and has not been peer reviewed. Data may be preliminary. n hw opredapo rgoi 2,2] t oei h rgeso n rgoi fO a been has the OS comparing of patients, prognosis OS and 54 PDGF- from progression and PDGF- specimens the BB and surgical in PDGF-AA examined role found - al. Its They (-AA, et 29]. PDGF Kubo of [28, 31]. level prognosis [30, tumors, solid poor well several though a as of metastasis targetable, investigated portend and was tumorigenesis to the IGF-1R shown in implicated that and been concluded tumors has factor authors unresectable growth derived The with Platelet patients months. OS 8.2 needed. with while of was patients OS months, investigation OS survival 6 20 In additional overall of was [27]. median 4 survival sarcoma al. a overall in Ewing 2 et had and phase growth median multi-institutional OS Kolb tumor the a relapsed delay conducted with tumors patients to resectable al. 21-25]. in et models robatumumab Anderson [18, using tumor study [26]. the xenograft antibodies clinical EFS inhibiting OS in inhibitory improvement by in significant invasion and with antibody, and xenografts anti-IGF-1R siRNA, migration an miRNA, preclinical proliferation, R1507, of cell using used number suppressed pathway A prognostic successfully signaling targets. have independent therapeutic IR/IGF-1R lines an as cell investigated be OS been have using to IGF-1R it and IR [20]. demonstrating Both samples associated analyses was OS expression Cox the IGF-1R multivariate within High higher with survival. bone marker. significantly to non-cancerous survival, correlated be versus was poorer OS to expression [19]. 26 found with IGF-1R IGF-1R in metastasis were samples, of proteins levels OS OS over-expression and 84 proteins the to that mRNA Using and showed IGF-1R in leading mRNA of pathway subsequently levels al. both signaling resistance, expression et samples; compared IR/IGF-1R drug Li the al. and 18]. et of survival, Wang [17, role proliferation, cancers the cellular various characterized promotes of have metastasis that and target, reports tumorigenesis feasible demonstrate numerous a to remains are needed. failed Her-2 is There results While value These Trastuzumab. clinical alone of respectively. its chemotherapy EFS into addition 50%, trastuzumab with investigation the 30-month and treated further and by 32% The survival overexpression chemotherapy of in Her-2 with improvement survival [16]. without significant overall treated Patients Her-2 and overexpression overexpressed respectively. EFS Her-2 demonstrated tumors 59%, with and whose patients Trastuzumab 96 32% patients involving of for were feasibility trial, in survival and clinical chemotherapy safety overall II the to phase xenograft and determine A adjunct to orthotopic sought tumors. an an OS, explanted as metastatic the within diagnosed in OS newly efficiency with in forming patients sarcosphere (TICs) utilized in cells al. reduction a et tumor-initiating Rainusso cellular target of [15]. abilities. inhibition to has migratory/invasive model time-dependent target of T-cells and inhibition dose- therapeutic CAR a and a found Her-2-specific rates, as They apoptotic role [14]. higher lines its proliferation, cell Her-2, OS standard of in utility phosphorylation, was prognostic was status pursued. expression the Her-2 Her-2 regarding been that concluding evidence demonstrate disease, conflicting to localized Despite unable OS with were diagnosed patients newly They in 1.10-3.71, with survival and patients CI: [13]. prognostic. overall 149 95% biopsy not Her-2 or of 2.02, both EFS of study = prospective value HR with in large and prognostic associated survival a 0.014; the overall conducted = determine COG p decreased the 1.16-3.72, to with Finally, CI: 95% al. 0.024). associated positive 2.07, = et Her-2 was [12]. = P being Zhang Her-2 (HR survival as contrast, specimens samples overall of removed identified In vs. and Overexpression and surgically (79.7% and outcomes respectively). expression survival events survival 1.09-5.67, negative. provided Her-2 adverse overall that and of between or studies and risk 1.07-4.72 relationship OS the 0.03) 16 CI, the increased identified = 95% evaluating They Her-2 p 2.54; meta-analysis of levels years, and a decreased 2.24 5 performed Additionally, at ratio: 0.03). with 45.6% = (rate associated p death was years, over-expression vs. 5 Her-2 (72.2% that at EFS found 58.2% They increased [11]. significantly chemotherapy both and surgery with treated OS nvivo In β nvitro In i o orlt oifro F p=01) hyas vlae mtnbmslt sa as mesylate imatinib evaluated also They 0.15). = (p EFS inferior to correlate not did diitaino h e--pcfi el infiatyrdcdTC,a vdne by evidenced as TICs, reduced significantly cells T Her-2-specific the of administration ,- α, tde yLn ta.ivsiae h oeo aaii,a niio fHer-2 of inhibitor an Lapatinib, of role the investigated al. et Long by studies B - BB, β eetrepeso hog muoitceityt ain prognosis. patient to immunohistochemistry through expression receptor ) α eetr eecreae ihifro F (p EFS inferior with correlated were receptors 4 < .5,wiePDGF- while 0.05), nvitro in studies Posted on Authorea 18 May 2020 — CC-BY 4.0 — https://doi.org/10.22541/au.158981322.20981629 — This a preprint and has not been peer reviewed. Data may be preliminary. u nig orbrt rvossraercpo xrsinrsls hyd o pert ffrobvious offer to While appear not offer. do could they receptors results, these patient- value expression of prognostic panel receptor what wide surface a establish previous across to receptors corroborate sought surface findings also cell We six our of samples. expression variable OS useful. the A derived validated increasingly we prove pathways. study, may intracellular this surfaceome In associated osteosarcoma’s of given expression independent a consistent of ideal, opportunities, preclinical are understanding targeting the patterns comprehensive offer receptor in or may explored receptors being overexpression expressed been currently uniquely and is has While conjugate, radioimmunotherapy been COG. antibody-drug the have targeted targeting by HER-2 which IGF2-R pursued a of OS. deruxtecan), the (trastuzumab using both of DS-8201 accomplished uses conjugates, and a setting be and arena antibody-drug can the pathway-independent as Targeting and in is receptor interest. radioimmunotherapy interest approach of including critical a of latter cell and means, the The using functional to of agent from be impact. variety therapeutic to pathway an a the pathway have directing associated the to for needs its solely is approach receptor and measure former The therapeutic receptor the cell. a expressing if inhibiting the targeting distinguish of +/- to means histologic 83.4 investigation.. the valuable and therapy, further is adjunctive 10% support tolerable It +/- not a 57.5 did is were standard EFS bevacizumab rate to while and survival that adjunct responses overall concluded an pre- tumor and They as both chemotherapy EFS bevacizumab respectively. free. and of 4-year %, bevacizumab progression 7.8 role estimated received were the OS The evaluate who improved localized post-operatively. to patients with everolimus patients and trials of and Thirty-one clinical proportion sorafenib months [47]. 2 small with treatment phase six a OS treatment completed at having that free al. despite demonstrate progression et months, despite to Navid were six progressed failed 17 at had They enrolled, progression that patients disease 28-61). anti- OS 38 an CI the high-grade sorafenib, 95% Of unresectable of (45%; efficacy with [46]. the patients treatment assessing chemotherapy in trial standard clinical everolimus 2 and phase antibody, non-randomized, a VEGF conducted [43-45]. al. lines proliferation, et cell cell previously Grignani suppressed OS the with successfully and standard to pathway associated in Similar VEGF the angiogenesis be [39-42]. both inhibit and target to to metastasis invasion miRNAs therapeutic and literature of a variety angiogenesis as a the of investigated utilized have promotion in been has its reported VEGF to receptors, extensively due discussed OS been in 6-month has prognosis had factor poor 20-50) CI growth 95% effects, endothelial (33%; anti-tumor Vascular patients demonstrated 42 and of tolerated 95% 14 well (12%; investigation. months; was patients further 6 42 cabozantinib warranting by of that Five cell responses concluded They Ewing [38]. objective OS advanced radiographically had non-progression. with decreased and OS patients histologically Sar- cabozantinib with in French both cabozantinib 4-26) The that efficacy using CI assess [37]. showed trial to microenvironment clinical OS, their combined or al. and 2 sarcoma cells phase et OS a Fioramonti conducted on investigated Group effects been coma its has settings. through c-Met, clinical migration of progres- and inhibitor and tumor proliferation an inhibit pre-clinical Cabozantinib, to in 36]. target [35, potential both invasion a and as migration investigated proliferation, been cell has transition metastasis. epithelial-mesenchymal c-Met the Thus, and in sion [34]. implicated cells and tumor described well of was been (EMT) imatinib has pathway showing to signaling in MET according children unsuccessful The response in were demonstrated imatinib They patients of treatment. OS (RECIST). effects single-agent Tumors their effective, the of Solid at an None in looking Criteria [33]. study Evaluation tumors clinical cell solid Response 2 inhibiting relapsed phase effects, evaluated or a synergistic refractory conducted al. yielded with COG therapy et combination Yamaguchi The that [32]. studies. demonstrated proliferation clinical They phase tumors. early OS and injected pre-clinical both in the utilized pathway been and has cytotoxicity Imatinib achieve unfeasible. to approach required therapeutic were doses this high making excessively inhibition, However, OS. for agent therapeutic nvivo in nitmreet fiaii essiaii n oouii nmc ihheterotopically with mice in doxorubicin and imatinib versus imatinib of effects anti-tumor nvitro In tde tlzn iN oihbtcMthv enscesu npreventing in successful been have c-Met inhibit to miRNA utilizing studies 5 nvitro in n lncly Studies clinically. and Posted on Authorea 18 May 2020 — CC-BY 4.0 — https://doi.org/10.22541/au.158981322.20981629 — This a preprint and has not been peer reviewed. Data may be preliminary. 3 olc,S,e al., study. et biology prospective S., Gorlick, studies. 13. 16 of meta-analysis and al., review et systematic Q., Zhang, 12. 2002. Cancer, al., et tumors. T., osteosarcoma Akatsuka, canine 11. in and models mouse in PDX al., et S., Karkare, 10. (IGF2R). receptor factor-2 growth al., like et D.S., Geller, 9. al., 740-9. et S.E., Hassan, 8. cohort. Study) Osteosarcoma American and (European al., EURAMOS-1 et S., Smeland, 7. al., et treatment. pre-randomisation J.S., Whelan, methotrexate. 6. high-dose and and doxorubicin, Italian the cisplatin, 2004-11. by to study tripeptide al., joint muramyl et P.A., a Meyers, extremity: 5. the of osteosarcoma localized Groups. with Sarcoma patients Scandinavian for doxorubicin al., and et Program. S., Results Ferrari, End 4. and Epidemiology, Surveillance, Savage, the S.A. and from Troisi, data R.J. L., Mirabello, 3. P., Kaatsch, 2. persons. elderly and Savage, ages S.A. and further middle Troisi, adolescents, and R.J. L., approaches Mirabello, targeted One, 1. future in cases. useful targeted chemo-resistant prove a and/or may References warranted. in metastatic, discussed is receptors receptors relapsed, strategies these such the all into of of that investigation address unlikely more combination, it adequately or a makes will burden with or one mutational treatment together using high taken single and of finding, variability a genomic This possibility demon- Osteosarcoma’s they samples. the utility, manner. OS highlights prognostic therapeutic patient-derived provide publications, of tumor. to panel prior whole fail a the receptors many may across of surface Findings representative expression cell variable heterogeneous. be six evaluated strated genomically not the may be we although Lastly, that to summary, known results results. 50 In is yielding statistical the that error, impact utilized tumor sampling We could larger by cutoff time. a limited over this from be stable varying sample is that small expression recognizing a that positivity, analyzed assuming for time, threshold the in a recapitulate point as adequately single not a does the at environment experimental tested raises The tumors factors. target. most several experience. human therapeutic by across limited a receptors is as study serve most This could of more expression or frequent one that the possibility Nevertheless, utility. prognostic 94 pdmooyo hlho cancer. childhood of Epidemiology 5:p 1397-404. p. (5): nvitro In eajvn hmteaywt ihds fsaie ihds ehteae cisplatin, methotrexate, high-dose Ifosfamide, high-dose with chemotherapy Neoadjuvant uvvladponsswt sesroa ucmsi oeta 00ptet nthe in patients 2000 than more in outcomes osteosarcoma: with prognosis and Survival agtdteayo sesroawt ailbldmncoa nioyt ninsulin- an to antibody monoclonal radiolabeled with osteosarcoma of therapy Targeted eeto n agtn nui rwhfco eetrtp IFR nosteosarcoma in (IGF2R) 2 type receptor factor growth insulin targeting and Detection E- xrsini ipyadsria pcmno rgoi fotoacm:A osteosarcoma: of prognosis on specimen surgical and biopsy in expression HER-2 elsraercpo xrsinpten nosteosarcoma. in patterns expression receptor surface Cell E- xrsini o rgotci sesroa hlrnsOclg Group Oncology Children’s a osteosarcoma; in prognostic not is expression HER-2 sesroa admzd rsetv ra fteadto fiofmd and/or ifosfamide of addition the of trial prospective randomized, a Osteosarcoma: rB xrsini orltdwt nrae uvvlo ainswt osteosarcoma. with patients of survival increased with correlated is expression ErbB2 eit lo acr 2014. Cancer, Blood Pediatr UAO-,a nentoa admsdsuyfrotoacm:rslsfrom results osteosarcoma: for study randomised international an EURAMOS-1, uosd o nieyrflc the reflect entirely not do tumors n no,2015. Oncol, Ann lnOcl 2005. Oncol, Clin J ulMdBo,2016. Biol, Med Nucl n acr 2009. Cancer, J Int sesroaicdneadsria ae rm17 o2004: to 1973 from rates survival and incidence Osteosarcoma nentoa sesroaicdnepten ncide and children in patterns incidence osteosarcoma International 26 acrTetRv 2010. Rev, Treat Cancer eiie(atmr) 2016. (Baltimore), Medicine 23 2:p 407-14. p. (2): 6 61 3) .8845-52. p. (34): 9:p 1558-64. p. (9): nvivo in c e,2019. Rep, Sci 43 1) .812-817. p. (12): 125 tt.Mroe,rcpo xrsinwas expression receptor Moreover, state. acr 2009. Cancer, u acr 2019. Cancer, J Eur 1:p 229-34. p. (1): 9 36 lnOcl 2005. Oncol, Clin J 1:p 11476. p. (1): 4:p 277-85. p. (4): 95 acr 2012. Cancer, 2) .e3661. p. (23): 115 7:p 1531-43. p. (7): 109 .36-50. p. : th 118 23 percentile 9:p. (9): 3:p. (3): Posted on Authorea 18 May 2020 — CC-BY 4.0 — https://doi.org/10.22541/au.158981322.20981629 — This a preprint and has not been peer reviewed. Data may be preliminary. o mtnbmslt hrp nosteosarcoma. in therapy mesylate imatinib al., for et T., Kubo, 31. osteosarcoma. al., in loop et paracrine and/or I., autocrine Sulzbacher, 30. outcome. adverse with associated 1520-7. al., is and et chondrosarcoma I., conventional Sulzbacher, 29. al., et cells. sarcoma A., Uren, 28. sarcoma. Ewing and osteosarcoma al., relapsed et with P.M., Anderson, 27. xenografts. osteosarcoma of growth inhibiting 55 in rapamycin with al., combination et E.A., Kolb, 26. osteosarcoma. al., human et in targets S., Avnet, 25. 2013. al., Cancer, et BMC M.L., Kuijjer, 24. IGFIR. Targeting by al., Chemosensitivity et Y., Liu, 23. al., et IGF-1R. G., Chen, (IGF-1R). 22. Receptor Factor Growth Insulin-Like Type-1 4132-4140. Targeting by al., coma et S.W., Zheng, osteosarcoma. 21. with patients in prognosis and al., metastasis et Y.H., Wang, 20. al., et 2019. Y.S., Li, 19. cancer. in IR and IGF-1R cotargeting 2652-64. for rationale (IGF-1R): receptor al., 1 et E., Buck, 18. 2012. Cancer, the from M., report Pollak, a 17. overexpression: 2 receptor factor growth group. epidermal oncology children’s human with osteosarcoma metastatic al., of et D., Ebb, 16. osteosarcoma. in cells al., initiating et N., vitro. Rainusso, in 15. axis HER2-PI3K/AKT-FASN the of activity al., the et X.H., Long, 14. 1:p 67-75. p. (1): 43 elPyilBohm 2016. Biochem, Physiol Cell 3:p 228-235. p. (3): 12 noee 2003. Oncogene, h nui n nui-iegot atrrcpo aiyi epai:a update. an neoplasia: in family receptor factor growth insulin-like and insulin The 3:p 159-69. p. (3): i-0 niisOtoacm elPoieain irto,adIvso n Enhances and Invasion and Migration, Proliferation, Cell Osteosarcoma Inhibits MiR-100 13 hs Itilo rsuua ncmiainwt yooi hmteayfrtreatment for chemotherapy cytotoxic with combination in trastuzumab of trial II Phase opnaoyislnrcpo I)atvto nihbto fislnlk rwhfactor- growth insulin-like of inhibition on activation (IR) receptor insulin Compensatory h osberl fislnlk rwhfco- nosteosarcoma. in factor-1 growth insulin-like of role possible The ltltdrvdgot atrrcpo saponsi akradateaetctarget therapeutic a and marker prognostic a as receptor factor growth Platelet-derived irRA13 niisOtoacm el rlfrto n nainvaTargeting via Invasion and Proliferation Cells Osteosarcoma Inhibits MicroRNA-133a eapaee-eie rwhfco eetrmdae oiiyadgot fEwing’s of growth and motility mediates receptor factor growth Beta-platelet-derived 10,aflyhmnmncoa nioytreigIF1,i ffcieaoeadin and alone effective is IGF-1R, targeting antibody monoclonal human fully a R1507, nui eetrioomAadislnlk rwhfco Ia diinltreatment additional as II factor growth insulin-like and A isoform receptor Insulin aaii lestemlgatpeoyeo sesroaclsvadwrglto of downregulation via cells osteosarcoma of phenotype malignant the alters Lapatinib .245. p. : nrae xrsino nui-iegot atr1rcpo screae ihtumor with correlated is receptor factor-1 growth insulin-like of expression Increased etcri upessPoieain irto,adIvso fHmnOsteosar- Human of Invasion and Migration, Proliferation, Suppresses Leptocarpin muohrp agtn E2wt eeial oie el lmntstumor- eliminates cells T modified genetically with HER2 targeting Immunotherapy RIFRsgaiga oeta agtfrteteto ihgaeosteosarcoma. high-grade of treatment for target potential as signaling IR/IGF1R lnOcl 2012. Oncol, Clin J ltltdrvdgot atraparcpo xrsinspot h rwhof growth the supports expression receptor factor-alpha growth Platelet-derived hs Isuyo lnclatvt fSH775 rbtmmb npatients in (robatumumab) 717454 SCH of activity clinical of study II phase A ltltdrvdgot atrA n aparcpo xrsinsget an suggests expression receptor -alpha and factor-AA growth Platelet-derived 22 acrRs 2009. Res, Cancer acrGn hr 2012. Ther, Gene Cancer 1) .2334-42. p. (15): 38 eho acrRsTet 2016. Treat, Res Cancer Technol 2:p 598-608. p. (2): 30 acr 2008. Cancer, eit lo acr 2016. Cancer, Blood Pediatr 2) .2545-51. p. (20): o ahl 2000. Pathol, Mod 69 7 6:p 2443-52. p. (6): ugOcl 2012. Oncol, Surg J no e,2014. Rep, Oncol 19 3:p 212-7. p. (3): 112 1) .2119-29. p. (10): 13 mJSr ahl 2001. Pathol, Surg J Am 15 o acrTe,2010. Ther, Cancer Mol 6:p 632-7. p. (6): 5:p NP40-8. p. (5): 105 31 e c oi,2017. Monit, Sci Med 63 eit lo acr 2010. Cancer, Blood Pediatr 1:p 328-34. p. (1): 3:p 235-43. p. (3): 1) .1761-70. p. (10): urPolCancer, Probl Curr 25 9 a Rev Nat 23 1) p. (12): 1) p. (10): p. : Posted on Authorea 18 May 2020 — CC-BY 4.0 — https://doi.org/10.22541/au.158981322.20981629 — This a preprint and has not been peer reviewed. Data may be preliminary. al 5. Table receptors 4. Table 3. Table 2. Table 1. Table Legend therapy. al., et F., Navid, 47. trial. clinical 2 phase non-randomised 98-107. a al., treatment: standard et after progressing G., Grignani, 46. al., pathway. et VEGFA/VEGFR1 L., Zhang, 45. osteosarcoma. in al., invasion et T., Yan, 44. VEGFA/Rictor. targeting by al., et T., Lv, 43. osteosarcoma. al., of prognosis et and Osteosarcoma. G., in Han, Prognosis Poor 42. and Toni, Metastasis N.D.M. Lung and for Khalil, Predictor E.Z.I. a F.E.A., Mohamed, 41. al., coma. et Y., Liu, 40. Perez-Soler, R. and S.E. blockade. Abdullah, 39. trial. 2 phase single-arm, multicentre, al., a et A., Italiano, 38. Microenvironment. Bone In Modifications al., and et Cells M., Fioramonti, 37. osteosarcoma. al., et in W., Xie, 36. c-Met. al., Targeting et Q., Li, 35. Lee, J. 2017. and Med, H.M. Transl Jeon, 34. study. Group Oncology al., Children’s et a M., tumors: Bond, 33. cells. osteosarcoma receptor-expressing factor al., growth et S.I., Yamaguchi, 32. ahlRsPat 2017. Pract, Res Pathol n acr 2017. Cancer, J Int acr 2012. Cancer, a lwCtmtyEpeso aaepesdi emti enFursetItniy(MFI). Intensity Fluorescent Mean Geometric in expressed Data Expression Cytometry Flow Raw ubro vrl uvvleet e ain ihrsett oiieo eaiereceptors negative or positive to respect with patient per events Survival Overall of Number receptor. each for negative or positive are that lines Cell (Autofluorescence) Antibodies Isotype Respective Their and Antibodies of Summary ubro vn reSria ES vnsprptetwt epc opstv rnegative or positive to respect with patient per events (EFS) Survival Free Event of Number i-0 sdw-euae notoacm n upesdM6 rlfrto n invasion and proliferation MG63 suppressed and osteosarcoma in down-regulated is miR-503 elMlMd 2019. Med, Mol Cell J irRA49-pSprse rlfrto,Mgain n naino sesroaby Osteosarcoma of Invasion and Migration, Proliferation, Suppresses MicroRNA-449b-5p e c oi,2019. Monit, Sci Med ihepeso eeso y6 n EFaeascae ihpo rgoi nosteosar- in prognosis poor with associated are VEGF and Cyr61 of levels expression High 5 irRA4 agt aclredteilgot atrt nii elpoieainand proliferation cell inhibit to factor growth endothelial vascular targets MicroRNA944 irRA865 niiscl rlfrto,mgainadivso ytreigc-Met targeting by invasion and migration proliferation, cell inhibits MicroRNA-876-5p hs Itileautn h esblt fadn eaiua osadr osteosarcoma standard to bevacizumab adding of feasibility the evaluating trial II phase A ffcso aclredteilgot atrepeso nptooia characteristics pathological on expression factor growth endothelial vascular of Effects 1:p 5. p. (1): irRA14ihbt sesroaagoeei n rlfrto ytreigthe targeting by proliferation and angiogenesis osteosarcoma inhibits MicroRNA-134 hs Isuyo mtnbmslt ncide ihrfatr rrlpe solid relapsed or refractory with children in mesylate imatinib of study II phase A aoatnbi ainswt dacdEigsroao sesroa(CABONE): osteosarcoma or sarcoma Ewing advanced with patients in Cabozantinib 118 oaei n vrlmsfrptet ihurscal ihgaeosteosarcoma high-grade unresectable with patients for everolimus and Sorafenib o e e,2018. Rep, Med Mol yegsi nirlfrtv ffc fiaii n dimcni platelet-derived in adriamycin and imatinib of effect antiproliferative Synergistic aoatnbAet sesroaGot hog ietEetO Tumor On Effect Direct A Through Growth Osteosarcoma Affects Cabozantinib ESJ 2018. J, FEBS 1) .3455-67. p. (14): acrBoak 2018. Biomark, Cancer 141 E:rlsi pteilmsnhmltasto n acrstemness. cancer and transition epithelial-mesenchymal in roles MET: 213 lnEpMd 2016. Med, Exp Clin 7:p 1469-1477. p. (7): 8:p 895-899. p. (8): 25 actOcl 2020. Oncol, Lancet 23 ehnsso eitnet aclredteilgot factor growth endothelial vascular to resistance of Mechanisms 285 .6236-6243. p. : 5:p 3293-3301. p. (5): eit lo acr 2008. Cancer, Blood Pediatr 18 7:p 1359-1371. p. (7): 6:p 5221-5228. p. (6): 23 c e,2018. Rep, Sci 8 acrSi 2015. Sci, Cancer 16 3:p 315-322. p. (3): aeln1Epeso oehrwt EFcnbe can VEGF with Together Expression Caveolin-1 4:p 577-584. p. (4): 21 3:p 446-455. p. (3): 8 1:p 4177. p. (1): 106 ahlOclRs 2019. Res, Oncol Pathol 50 7:p 875-82. p. (7): actOcl 2015. Oncol, Lancet 2:p 254-8. p. (2): 16 1:p. (1): Ann Posted on Authorea 18 May 2020 — CC-BY 4.0 — https://doi.org/10.22541/au.158981322.20981629 — This a preprint and has not been peer reviewed. Data may be preliminary. ufc receptors. surface geoMFI 1. Table group Supplementary pattern expression high (C) 3. group Figure pattern expression medium (B) group pattern 2. Figure control negative corresponding 1. Figure 7. Table 6. Table positive aadrtoaayi o vrl uvvlprcl ufc receptor surface cell per survival overall for receptor analysis surface ratio cell Hazard per EFS for analysis ratio Hazard eetrepeso atrsasge ygoercma ursetitniy()lwexpression low (A) intensity fluorescent mean geometric by assigned patterns expression Receptor oa ubro oiiercposprcl ieuigte50 the using line cell per receptors positive of number Total odcag nrcpo xrsindfie srltv nraei xrsinrltv othe to relative expression in increase relative as defined expression receptor in change Fold /geoMFI negative Monoclonal anti-human (APC) VEGFR-3 Monoclonal anti-human HGFR/c-MET (APC) Polyclonal anti-human IR(AF488) Monoclonal anti-human (FITC) IGF-1R Monoclonal anti-human CD140b/PDGFR- Monoclonal anti-human HER-2/neu (PE) Table 1. Summary of Antibodies and Their Respective Isotype Antibodies (Autofluorescence) Receptor Antibody (Fluorophore) oprsnbtenHsa ta.adtecretsuy eosrtn mean demonstrating study, current the and al. et Hassan between Comparison n offiin fvrain(enda tnaddvainma)ars l six all across deviation/mean) standard as (defined variation of coefficient and β (PE) R&D/FAB3492A R&D/FAB3582A R&D/FAB1544G R&D/FAB391F R&D/FAB1263P BD/340552 Vendor/Catalog No. 9 IgG/NP_0010732 mouse mouse goat mouse mouse mouse IgG1/54733 IgG1/95106 IgG1/33255 IgG1/PR7212 IgG1/Neu24.7 85 Isotype Antibody monocytes (periph blood monocytes (periph blood monocytes (periph blood MCF7 ATCC/HTB-22) (ATCC/CRL- 697.Sp Hs (ATCC/HTB-22)MCF7 7433) sample) sample) sample) th ecniea uo o positivity. for cutoff a as percentile Positive Control Posted on Authorea 18 May 2020 — CC-BY 4.0 — https://doi.org/10.22541/au.158981322.20981629 — This a preprint and has not been peer reviewed. Data may be preliminary. C396 C379 C377 C375 C374 C373 C371 C370 C368 C366 C365 C360 C353 C349 C346 C342 C340 C338 C337 C334 C331 C327 C326 C323 C315 C314 C311 C307 C301 C300 C299 C297 C293 C291 C290 C287 C282 C281 C256 C253 C251 C244 C243 C238 C233 C232 C231 C219 C209 C205 C204 C194 Cell Line Table 2. Raw Flow Cytometry RawFlow Expression Data expressed in Geometric Mean Fluorescent Intensity (MFI). 18102 13516 10606 10605 10890 4040 3154 2256 7012 5245 4736 6878 2955 3640 5244 2148 3441 5275 5374 2920 3356 2161 7755 4273 8697 3443 1483 2682 5177 2451 3125 4357 4020 3160 3997 3532 2225 6446 4661 2345 4378 7002 2446 1731 3391 3968 2925 4281 3920 4307 2436 3813 GM HER-2 Negative 1462 1276 1473 1431 1229 GM 384 252 435 548 379 637 137 612 846 430 605 610 348 269 842 386 487 353 882 304 859 521 198 305 569 129 337 263 394 375 445 375 377 553 552 276 425 236 231 301 128 237 722 376 569 187 274 44596 10890 23437 12751 13316 12818 10191 22774 26729 6416 9757 9037 1230 2066 6503 5408 5284 1248 3298 1285 4210 5369 4091 1901 4561 1674 1372 2035 1401 1904 4867 4540 5033 3198 7153 5676 2370 3653 2755 3028 2743 4741 4012 4475 2272 5361 4001 1177 5113 GM 729 941 652 PDGFR- Negative β 1431 1229 1462 1276 1473 GM 384 252 435 548 379 637 337 612 846 430 605 610 348 269 842 386 487 353 882 304 859 521 198 305 569 129 337 263 394 375 445 375 377 553 552 276 425 236 231 301 128 237 722 376 569 187 274 1260 GM 966 447 319 639 843 462 271 442 228 402 546 476 740 739 331 580 865 616 475 490 360 586 391 988 604 289 353 685 177 351 379 396 526 398 464 469 749 734 307 445 368 349 474 404 431 461 575 338 982 195 366 IGF-1R Negative 10 GM 351 351 351 351 351 351 351 351 351 351 351 351 271 351 351 351 351 351 351 351 351 340 351 351 351 351 351 351 351 351 351 351 351 351 351 351 351 351 351 351 351 351 351 351 351 351 351 351 351 351 351 351 1378 1061 1076 1148 GM 355 453 577 571 426 498 613 766 349 685 498 437 522 392 446 590 556 474 212 417 480 380 695 769 455 733 272 468 607 537 900 816 398 601 900 724 486 534 413 704 363 843 606 326 443 566 210 409 IR Negative GM 442 442 442 442 442 442 442 442 442 442 442 442 442 442 442 442 442 442 442 442 442 442 442 442 442 442 442 442 442 442 442 442 442 442 442 442 442 442 442 442 442 442 271 442 442 442 442 442 442 442 442 431 2420 3957 2775 2934 3038 3170 1523 1031 1802 3107 5352 1688 1334 8388 1019 2565 1356 6160 1902 1358 1816 1915 1845 1079 2534 1570 3677 3047 1176 1170 1134 1146 1196 1112 GM 835 904 957 726 821 828 670 593 531 990 994 830 903 993 812 752 844 315 c-MET Negative GM 830 830 830 830 830 830 830 830 830 830 830 830 529 830 830 830 830 830 830 830 830 808 830 830 830 830 830 830 830 830 830 830 830 830 830 830 830 442 830 830 830 830 830 830 830 830 830 830 830 830 830 830 1046 2224 1403 1299 1028 1491 1000 1012 GM 378 464 436 726 553 581 891 361 764 547 517 690 664 513 618 864 466 208 444 393 264 417 551 419 701 226 627 737 560 659 836 395 237 690 567 558 406 366 621 223 419 725 177 416 VEGFR-3 Negative GM 520 520 520 520 520 520 520 520 520 520 520 520 520 520 520 520 520 520 520 520 520 520 520 520 520 520 520 520 520 520 520 520 520 520 520 520 520 520 520 520 520 520 529 520 520 520 520 520 520 520 520 507 Posted on Authorea 18 May 2020 — CC-BY 4.0 — https://doi.org/10.22541/au.158981322.20981629 — This a preprint and has not been peer reviewed. Data may be preliminary. Table 3. Cell line C396 C379 C377 C375 C374 C373 C371 C370 C368 C366 C365 C360 C353 C349 C346 C342 C340 C338 C337 C334 C331 C327 C326 C323 C315 C314 C307 C301 C300 C299 C297 C293 C291 C290 C287 C282 C281 C256 C253 C251 C244 C243 C238 C233 C232 C231 C219 C209 C205 C204 C194 C311 Total Cell lines that are positive ornegative foreach receptor. negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive HER-2 26 PDGFR- negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive 26 β negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive IGF-1R 26 negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive IR 25 11 negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive c-MET 26 VEGFR-3 negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative negative positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive positive 26 Total 4 3 3 5 4 1 5 4 2 0 4 4 2 2 6 1 4 2 2 6 2 2 3 6 5 2 5 2 2 5 4 3 5 0 5 4 3 3 1 0 4 2 5 4 0 1 5 1 2 1 2 2 Posted on Authorea 18 May 2020 — CC-BY 4.0 — https://doi.org/10.22541/au.158981322.20981629 — This a preprint and has not been peer reviewed. Data may be preliminary. positiv Table 4. e orreceptorsnegativee

Number of E Number of vent F ree

S urvival urvival (EFS) 12

events events per with per patientrespect to

Posted on Authorea 18 May 2020 — CC-BY 4.0 — https://doi.org/10.22541/au.158981322.20981629 — This a preprint and has not been peer reviewed. Data may be preliminary.

negative receptors T

able 5.

Number of Overall Survival events per patient with respect to positive or

13

Posted on Authorea 18 May 2020 — CC-BY 4.0 — https://doi.org/10.22541/au.158981322.20981629 — This a preprint and has not been peer reviewed. Data may be preliminary. Table 6.

Hazard ratio per EFS cell analysis for receptor surface 14

Posted on Authorea 18 May 2020 — CC-BY 4.0 — https://doi.org/10.22541/au.158981322.20981629 — This a preprint and has not been peer reviewed. Data may be preliminary. Table 7.

Hazard ratio overall analysis for survival per cell 15

surface receptorsurface

Posted on Authorea 18 May 2020 — CC-BY 4.0 — https://doi.org/10.22541/au.158981322.20981629 — This a preprint and has not been peer reviewed. Data may be preliminary. relative to the corresponding Figure 1.

Fold Change Compared to Negative Control Fold change in receptor expression defined asrelative increase in expression Fold ChangeinReceptorExpressionRelativetoNegativeControl 40 80 10 0 HER-2 PDGFR

negative IGF-1R control Receptor 16

IR c-MET VEGFR-3

Posted on Authorea 18 May 2020 — CC-BY 4.0 — https://doi.org/10.22541/au.158981322.20981629 — This a preprint and has not been peer reviewed. Data may be preliminary. expression pattern group lowexp(A) Figure 2. (A) Geometric Mean Geometric Mean 1000 1500 1000 1500

500 500 0 0 C194 C194 C204

C204 Receptor expression patterns assigned bygeometric mean C205 C205

C209 C209 ression pattern group(B)medium expression pattern group(C)high C219 C219 C231 C231 C232 C232 C233 C233 C238 C238 C243 C243 C244 C244

C251 C251 C253 C253 C256 C256 C281 C281 C282 C282 C287 C287 C290 C290 C291 C291 C293 C293 C297 C297 C299 C299 17 C300 C300 C301 C301 C307 Cell lines C307 IGF-1R Cell lines C311 C311 C314

C314 IR C315 C315 C323 C323 C326 C326 C327 C327 C331 C331 C334 C334 C337 C337 C338 C338 C340 C340 C342 C342 C346

C346 fluorescent intensity C349 C349 C353 C353 C360 C360 C365 C365 C366 C366 C368 C368 C370 C370 C371 C371 C373 C373 C374 C374 C375 C375 C377 C377

C379 C379 C396 C396

Posted on Authorea 18 May 2020 — CC-BY 4.0 — https://doi.org/10.22541/au.158981322.20981629 — This a preprint and has not been peer reviewed. Data may be preliminary. positivity for cutoff Figure 3.

Total p number of Number of Cell Lines 10 12 14 0 2 4 6 8 . 0

Positive ReceptorsperCellLine 0/6 ositive receptors per cell line 50th PercentileCutoff 1/6 2/6 18 3/6 4/6 using 5/6

the 50 6/6 th

percentile

as a Posted on Authorea 18 May 2020 — CC-BY 4.0 — https://doi.org/10.22541/au.158981322.20981629 — This a preprint and has not been peer reviewed. Data may be preliminary.

PDGFRB VEGFR3 IGF1R cMET Her2 IR

standard deviation/mean) demonstrating mean geoMFI TableSupplementary 1.

geoMFI positive (Std. 0.43 (0.34) 0.67 (0.59) 1.69 (1.85) 0.26 (0.30) 3.42 (3.76) 0.31 (0.18) Mean

/geoMFI Dev Hassan et al.Hassan

. )

across all receptors. sixsurface

Comparison between et Hassan al. and the current study negative positive /geoMFI

Coefficient of Variation negative 0.79 0.88 1.09 1.19 1.09 0.59 19

and variation coefficient of

geoMFI

12.56 (12.19) 10.76 (6.06) positive (Std. 1.23 (0.71) 2.24 (1.81) 1.32 (0.54) 1.47 (0.63) Mean

/geoMFI Dev Cu

. rrent Study )

( def negative ined as ,

Coefficient of Variation 0.57 0.81 0.41 0.43 0.97 0.56