The Prognostic Importance of ctDNA in Rectal Cancer: Comparison
Please note this is a comparison between Version 2 by Beatrix Zheng and Version 1 by Edina Dizdarevic Lauridsen.

An individualized treatment approach is necessary to improve survival and quality of life in rectal cancer. Tools to stratify patients are currently missing. Circulating tumor DNA seems to be a good candidate, but results are sparse, conflicting and characterized by lack of a uniform approach. As tThe interpretation of results is dependent on the quality of reporting, we aimed to address this issue in our paper. Our results inctDNA status may be of importance to the long-term prognosis, but the area of research is new and is short of dedicate an association between ctDNA status and outcome. In six out of nine papers, bias was low. However, a conclusion is dubious because of the heterogeneity among the studies and lack of standardized methods. Studiesd studies. There is an obvious need for standardization in ctDNA research, and the issue should be addressing these issues are warranteded in future research

  • rectal cancer
  • colorectal cancer
  • ctDNA
  • liquid biopsy
  • biomarkers
  • QUIPS

1. Introduction

The treatment of locally advanced rectal cancer (LARC) has evolved from surgery alone to experimental trimodality treatment comprising neoadjuvant chemoradiotherapy (nCRT) followed by surgery. Modifications of radiation doses and treatment schedules have been presented with different rates of pathological response but without improvement of long-term survival [1][2][3][4][5]. Complete pathological response remains an important prognostic factor but is a poor surrogate endpoint for overall survival [6].
The achievement of complete pathological response after chemoradiation suggested the possibility of organ preservation. A selected group of rectal cancer patients achieving complete tumor regression, termed clinical complete response (cCR) have been treated according to this approach [7]. Several studies have reported excellent oncological outcome and high quality of life [8][9][10][11], but a relatively high fraction will recur during follow-up. It remains unclear how patients should be selected for surgery versus observation after chemoradiation. Total neoadjuvant therapy is a new approach in rectal cancer, but the concept needs further validation before it comes to clinical utility [12][13].
The treatment of rectal cancer calls for new markers, which alone or in combination can guide a more individualized treatment approach.
Circulating tumor DNA (ctDNA) is a new biomarker that has gained rapidly increasing interest. In many aspects, it seems to fulfill the criteria of an ideal tumor marker. It is easily accessible for repeated measurement with little discomfort to the patient, reflects important biological characteristics and seems to overcome the issue of tumor heterogeneity [14][15]. There is a steep increase in the number of publications, witnessing a considerable clinical interest. In colon cancer, several prospective trials are investigating if ctDNA can identify patients with non-metastatic colon cancer, with high risk of recurrence, that may benefit from adjuvant chemotherapy [16][17][18][19]. However, in rectal cancer, the influence of ctDNA on treatment decisions is still negligible. The diverging results and lack of standardization pose a major problem and impede further development. The situation calls for a comprehensive evaluation of the current literature.

2. Current Insights

The current literature points toward ctDNA as a promising tumor marker in clinical oncology. In rectal cancer, ctDNA may result in an individualized treatment approach. Reviews published during the recent years have tried to outline the role of ctDNA in rectal cancer, but none of them have addressed the quality of the included studies [20][21]. To our knowledge, our review is the first to address this aspect.
Bias was assessed with the QUIPS tool. Six studies were classified as having low bias, two with moderate risk and one with high risk of bias. The developers of the QUIPS suggest that studies with high risk of bias should be omitted from further analysis. Based on this statement, only six out of nine studies in ourthe reviewsearch would be eligible for the evaluation of ctDNA. Since the population in our reviewthe research is small and ourthe results are reported narratively, wethe researchers present all results from the analysis. However, this serves as a clear indicator of the necessity of a quality assessment for future reviews.
There is no evaluation of bias in papers reviewing ctDNA in rectal cancer. The lack of quality appraisal is a problem in many reviews. A literature search of Hayden et al. revealed a rising number of published reviews and a consistent problem with the quality assessment. Almost 50% of the articles generated from their search were excluded because of lacking quality assessment. For the remaining articles, the evaluation varied and was often inadequate. Because of this gap, they developed QUIPS to assess validity and bias in studies investigating prognostic factors [22]. This is the only tool specifically developed for quality evaluation of prognostic studies. Although the tool indicates potential problems with only three out of nine studies, it should be used cautiously, as it does not address all problems, i.e., comparison of methodological issues.
Riley et al. suggest a guide that should be applied when conducting systematic reviews and meta-analyses of prognostic studies [23]. In the future, this systematic approach will lead to fewer reviews with comprehensive analysis of the included literature. As a result, the role of different biomarkers may be clarified faster, and unnecessary research can be omitted.
The REMARK checklist was developed to ensure consistent reporting [24][25]. In spite of the introduction of the checklist, results have been discouraging with no improvement in reporting [26]. This is supported by outhe researchers' findings with only one study reporting the use of the checklist. Hence, there is a risk of continuous research in markers without real progress as to clinical utility.
Another issue is the lack of standardized methods for ctDNA detection and reporting, as this impairs comparison and generalization of results. This is indeed what wthe researchers see in this reviewentry. Characteristics and features of the tumor, which are not fully understood, can aggravate this problem along with other problems as illustrated in the literature [27][28][29][30][31][32][33]. The discussion of the methodology is outside the scope of this entreviewy, but a comprehensive reviewentry has addressed the current challenges and suggests possible solutions which should be considered in future research [34].
The predictive value of ctDNA varied among the included studies. Detectable ctDNA at baseline does not seem to be a predictor of treatment response. Only two of seven studies report the presence of ctDNA after CRT and a poor response as evaluated by MR or the surgical specimen, respectively [29][35]. These results indicate that ctDNA currently cannot be used to select treatment, i.e., organ preservation or surgery. In general, the results should be carefully considered, since none of the studies report power calculations. This is likely a result of the retrospective restudyearch design. For the prospective studies, the ctDNA analyses are exploratory, and it may be rather difficult to make proper assumptions regarding the power calculations. Post hoc power calculations are generally not recommended [36]. Hence, the literature is short of dedicated prospective trials exploring the clinical aspects of ctDNA in rectal cancer. However, in colon cancer, several prospective trials are investigating if ctDNA can predict recurrence and if ctDNA-guided treatment can improve survival [17][18][34][37]. Circulating tumor DNA seems promising in other types of cancers, but results are limited by the relatively small size of the trials [38][39][40].
For the survival outcomes, all of the studies found a correlation between survival outcome and ctDNA at a certain time point. For the baseline measurements, three out of nine reported an association between detectable ctDNA and worse survival. For the pre- and post-surgery analyses, four out of six and five out of five, respectively, found an association between detectable ctDNA and worse outcome. The persistence of ctDNA throughout the course of treatment seems to be a poor prognostic marker. The reported increasing HR at each time point supports this hypothesis. Especially detectable postoperative ctDNA seems to correlate with relapse. Similar results are reported in the treatment of colon cancer and malignant melanoma [38][41].
At this point, it is difficult to conclude as to the prognostic importance of ctDNA in rectal cancer as most studies are small, the statistical assumptions are dubious and the follow-up period is rather short in some of the included studies. There is a substantial risk of both missing a correlation and finding one that would not be present if a longer follow-up period were available. In a review assessing trial extensions using routinely collected data, the authors explored if there were any beneficial or harmful events in the period beyond the original trial time period. They showed that in nearly 50% of the trials, a benefit was observed with extended follow-up, and in nearly 30% of these, the benefits were only significant for this period. Loss of significant benefits was only seen in 7% of the studies [42]. These results highlight the importance of a sufficiently long follow-up period in terms of evaluating interventions, new diagnostic or prognostic tools. For rectal cancer, most local regrowths appear within the first two years, while metastatic disease may appear several years later. Therefore, both the follow-up period and the evaluation of the endpoint should be selected carefully prior to trial initiation [43]. Studies with large cohorts and long follow-up are time consuming and expensive, but they are necessary in terms of clarifying the real value of biomarkers. A surrogate endpoint would be ideal to select optimal treatment and reduce costs and follow-up in rectal cancer. The study published by Jakobsen et al. indicates that ctDNA response may serve as a surrogate marker for OS in several cancers [44]. Organ preservation in rectal cancer as standard treatment could be a possibility if ctDNA response proved to be a reliable surrogate marker for oncological outcome.

References

  1. Bosset, J.-F.; Collette, L.; Calais, G.; Mineur, L.; Maingon, P.; Radosevic-Jelic, L.; Daban, A.; Bardet, E.; Beny, A.; Ollier, J.-C. Chemotherapy with Preoperative Radiotherapy in Rectal Cancer. N. Engl. J. Med. 2006, 355, 1114–1123.
  2. van Gijn, W.; Marijnen, C.A.; Nagtegaal, I.D.; Kranenbarg, E.M.-K.; Putter, H.; Wiggers, T.; Rutten, H.J.; Påhlman, L.; Glimelius, B.; van de Velde, C.J.; et al. Preoperative radiotherapy combined with total mesorectal excision for resectable rectal cancer: 12-year follow-up of the multicentre, randomised controlled TME trial. Lancet Oncol. 2011, 12, 575–582.
  3. Sauer, R.; Liersch, T.; Merkel, S.; Fietkau, R.; Hohenberger, W.; Hess, C.; Becker, H.; Raab, H.-R.; Villanueva, M.-T.; Witzigmann, H.; et al. Preoperative Versus Postoperative Chemoradiotherapy for Locally Advanced Rectal Cancer: Results of the German CAO/ARO/AIO-94 Randomized Phase III Trial After a Median Follow-Up of 11 Years. J. Clin. Oncol. 2012, 30, 1926–1933.
  4. Rödel, C.; Graeven, U.; Fietkau, R.; Hohenberger, W.; Hothorn, T.; Arnold, D.; Hofheinz, R.-D.; Ghadimi, M.; Wolff, H.A.; Lang-Welzenbach, M.; et al. Oxaliplatin added to fluorouracil-based preoperative chemoradiotherapy and postoperative chemotherapy of locally advanced rectal cancer (the German CAO/ARO/AIO-04 study): Final results of the multicentre, open-label, randomised, phase 3 trial. Lancet Oncol. 2015, 16, 979–989.
  5. Julião, G.P.S.; Habr-Gama, A.; Vailati, B.B.; Aguilar, P.B.; Sabbaga, J.; Araújo, S.E.A.; Mattacheo, A.; Alexandre, F.; Fernandez, L.M.; Gomes, D.B.; et al. Is neoadjuvant chemoradiation with dose-escalation and consolidation chemotherapy sufficient to increase surgery-free and distant metastases-free survival in baseline cT3 rectal cancer? Eur. J. Surg. Oncol. (EJSO) 2018, 44, 93–99.
  6. Petrelli, F.; Borgonovo, K.; Cabiddu, M.; Ghilardi, M.; Lonati, V.; Barni, S. Pathologic complete response and disease-free survival are not surrogate endpoints for 5-year survival in rectal cancer: An analysis of 22 randomized trials. J. Gastrointest. Oncol. 2017, 8, 39–48.
  7. Habr-Gama, A.; Perez, R.O.; Nadalin, W.; Sabbaga, J.; Ribeiro, U., Jr.; Silva e Sousa, A.H., Jr.; Campos, F.G.; Kiss, D.R.; Gama-Rodrigues, J. Operative Versus Nonoperative Treatment for Stage 0 Distal Rectal Cancer Following Chemoradiation Therapy: Long-term results. Ann. Surg. 2004, 240, 711–718.
  8. van der Valk, M.J.M.; Hilling, D.; Bastiaannet, E.; Kranenbarg, E.M.-K.; Beets, G.L.; Figueiredo, N.; Habr-Gama, A.; O Perez, R.; Renehan, A.G.; van de Velde, C.J.H.; et al. Long-term outcomes of clinical complete responders after neoadjuvant treatment for rectal cancer in the International Watch & Wait Database (IWWD): An international multicentre registry study. Lancet 2018, 391, 2537–2545.
  9. Fernandez, L.M.; Julião, G.P.S.; Figueiredo, N.L.; Beets, G.L.; van der Valk, M.J.M.; Bahadoer, R.R.; E Hilling, D.; Kranenbarg, E.M.-K.; Roodvoets, A.G.H.; Renehan, A.G.; et al. Conditional recurrence-free survival of clinical complete responders managed by watch and wait after neoadjuvant chemoradiotherapy for rectal cancer in the International Watch & Wait Database: A retrospective, international, multicentre registry study. Lancet Oncol. 2021, 22, 43–50.
  10. Jones, H.J.; Al-Najami, I.; Cunningham, C. Quality of life after rectal-preserving treatment of rectal cancer. Eur. J. Surg. Oncol. (EJSO) 2020, 46, 2050–2056.
  11. Dizdarevic, E.; Hansen, T.; Pløen, J.; Jensen, L.H.; Lindebjerg, J.; Rafaelsen, S.; Jakobsen, A.; Appelt, A.L. Long-Term Patient-Reported Outcomes After High-Dose Chemoradiation Therapy for Nonsurgical Management of Distal Rectal Cancer. Int. J. Radiat. Oncol. 2020, 106, 556–563.
  12. Garcia-Aguilar, J.; Chow, O.S.; Smith, D.D.; Marcet, J.E.; Cataldo, P.A.; Varma, M.G.; Kumar, A.S.; Oommen, S.; Coutsoftides, T.; Hunt, S.R.; et al. Effect of adding mFOLFOX6 after neoadjuvant chemoradiation in locally advanced rectal cancer: A multicentre, phase 2 trial. Lancet Oncol. 2015, 16, 957–966.
  13. Fokas, E.; Schlenska-Lange, A.; Polat, B.; Klautke, G.; Grabenbauer, G.G.; Fietkau, R.; Kuhnt, T.; Staib, L.; Brunner, T.; Grosu, A.-L.; et al. Chemoradiotherapy plus Induction or Consolidation Chemotherapy as Total Neoadjuvant Therapy for Patients with Locally Advanced Rectal Cancer: Long-term Results of the CAO/ARO/AIO-12 Randomized Clinical Trial. JAMA Oncol. 2022, 8, e215445.
  14. Keller, L.; Belloum, Y.; Wikman, H.; Pantel, K. Clinical relevance of blood-based ctDNA analysis: Mutation detection and beyond. Br. J. Cancer 2021, 124, 345–358.
  15. Cescon, D.W.; Bratman, S.V.; Chan, S.M.; Siu, L.L. Circulating tumor DNA and liquid biopsy in oncology. Nat. Cancer 2020, 1, 276–290.
  16. Tie, J.; Cohen, J.D.; Wang, Y.; Christie, M.; Simons, K.; Lee, M.; Wong, R.; Kosmider, S.; Ananda, S.; McKendrick, J.; et al. Circulating Tumor DNA Analyses as Markers of Recurrence Risk and Benefit of Adjuvant Therapy for Stage III Colon Cancer. JAMA Oncol. 2019, 5, 1710–1717.
  17. Folprecht, G.; Reinacher-Schick, A.; Weitz, J.; Lugnier, C.; Kraeft, A.-L.; Wisser, S.; Aust, D.E.; Weiss, L.; von Bubnoff, N.; Kramer, M.; et al. The CIRCULATE Trial: Circulating Tumor DNA Based Decision for Adjuvant Treatment in Colon Cancer Stage II Evaluation (AIO-KRK-0217). Clin. Color. Cancer 2021.
  18. Taïeb, J.; Benhaim, L.; Puig, P.L.; Le Malicot, K.; Emile, J.F.; Geillon, F.; Tougeron, D.; Manfredi, S.; Chauvenet, M.; Taly, V.; et al. Decision for adjuvant treatment in stage II colon cancer based on circulating tumor DNA:The CIRCULATE-PRODIGE 70 trial. Dig. Liver Dis. 2020, 52, 730–733.
  19. Merk, C.; Martling, A.; Lindberg, J.; Benhaim, L.; Taieb, J.; Lind, P. Circulating tumor DNA (ctDNA) in adjuvant therapy of early stage colon cancer: Current status and future perspectives. Acta Oncol. 2022, 61, 523–530.
  20. Massihnia, D.; Pizzutilo, E.G.; Amatu, A.; Tosi, F.; Ghezzi, S.; Bencardino, K.; Di Masi, P.; Righetti, E.; Patelli, G.; Scaglione, F.; et al. Liquid biopsy for rectal cancer: A systematic review. Cancer Treat. Rev. 2019, 79, 101893.
  21. Morais, M.; Pinto, D.M.; Machado, J.C.; Carneiro, S. ctDNA on liquid biopsy for predicting response and prognosis in locally advanced rectal cancer: A systematic review. Eur. J. Surg. Oncol. (EJSO) 2021, 48, 218–227.
  22. Hayden, J.A.; Côté, P.; Bombardier, C. Evaluation of the Quality of Prognosis Studies in Systematic Reviews. Ann. Intern. Med. 2006, 144, 427–437.
  23. Riley, R.D.; Moons, K.G.M.; Snell, K.I.E.; Ensor, J.; Hooft, L.; Altman, D.G.; Hayden, J.; Collins, G.S.; Debray, T.P.A. A guide to systematic review and meta-analysis of prognostic factor studies. BMJ 2019, 364, k4597.
  24. McShane, L.M.; for the Statistics Subcommittee of the NCI-EORTC Working Group on Cancer Diagnostics; Altman, D.G.; Sauerbrei, W.; E Taube, S.; Gion, M.; Clark, G.M. REporting recommendations for tumour MARKer prognostic studies (REMARK). Br. J. Cancer 2005, 93, 387–391.
  25. Altman, D.G.; McShane, L.M.; Sauerbrei, W.; E Taube, S. Reporting recommendations for tumor marker prognostic studies (REMARK): Explanation and elaboration. BMC Med. 2012, 10, 51.
  26. Sekula, P.; Mallett, S.; Altman, U.G.; Sauerbrei, W. Did the reporting of prognostic studies of tumour markers improve since the introduction of REMARK guideline? A comparison of reporting in published articles. PLoS ONE 2017, 12, e0178531.
  27. Appelt, A.L.; Andersen, R.F.; Lindebjerg, J.; Jakobsen, A. Prognostic Value of Serum NPY Hypermethylation in Neoadjuvant Chemoradiotherapy for Rectal Cancer. Am. J. Clin. Oncol. 2020, 43, 9–13.
  28. Sclafani, F.; Chau, I.; Cunningham, D.; Hahne, J.C.; Vlachogiannis, G.; Eltahir, Z.; Lampis, A.; Braconi, C.; Kalaitzaki, E.; De Castro, D.G.; et al. KRAS and BRAF mutations in circulating tumour DNA from locally advanced rectal cancer. Sci. Rep. 2018, 8, 1445.
  29. Khakoo, S.; Carter, P.D.; Brown, G.; Valeri, N.; Picchia, S.; Bali, M.A.; Shaikh, R.; Jones, T.; Begum, R.; Rana, I.; et al. MRI Tumor Regression Grade and Circulating Tumor DNA as Complementary Tools to Assess Response and Guide Therapy Adaptation in Rectal Cancer. Clin. Cancer Res. 2020, 26, 183–192.
  30. Fiala, C.; Diamandis, E.P. Utility of circulating tumor DNA in cancer diagnostics with emphasis on early detection. BMC Med. 2018, 16, 1–10.
  31. Bettegowda, C.; Sausen, M.; Leary, R.J.; Kinde, I.; Wang, Y.; Agrawal, N.; Bartlett, B.R.; Wang, H.; Luber, B.; Alani, R.M.; et al. Detection of circulating tumor DNA in early- and late-stage human malignancies. Sci. Transl. Med. 2014, 6, 224.
  32. Lam, K.; Pan, K.; Linnekamp, J.F.; Medema, J.P.; Kandimalla, R. DNA methylation based biomarkers in colorectal cancer: A systematic review. Biochim. Biophys. Acta 2016, 1866, 106–120.
  33. Andersen, R.F. Tumor-specific methylations in circulating cell-free DNA as clinically applicable markers with potential to substitute mutational analyses. Expert Rev. Mol. Diagn. 2018, 18, 1011–1019.
  34. Dasari, A.; Morris, V.K.; Allegra, C.J.; Atreya, C.; Benson, A.B., 3rd; Boland, P.; Chung, K.; Copur, M.S.; Corcoran, R.B.; Deming, D.A.; et al. ctDNA applications and integration in colorectal cancer: An NCI Colon and Rectal–Anal Task Forces whitepaper. Nat. Rev. Clin. Oncol. 2020, 17, 757–770.
  35. Tie, J.; Cohen, J.D.; Wang, Y.; Li, L.; Christie, M.; Simons, K.; Elsaleh, H.; Kosmider, S.; Wong, R.; Yip, D.; et al. Serial circulating tumour DNA analysis during multimodality treatment of locally advanced rectal cancer: A prospective biomarker study. Gut 2019, 68, 663–671.
  36. Turner, D.P.; Houle, T.T. The Importance of Statistical Power Calculations. Headache J. Head Face Pain 2018, 58, 1187–1191.
  37. Nors, J.; Henriksen, T.V.; Gotschalck, K.A.; Juul, T.; Søgaard, J.; Iversen, L.H.; Andersen, C.L. IMPROVE-IT2: Implementing noninvasive circulating tumor DNA analysis to optimize the operative and postoperative treatment for patients with colorectal cancer – intervention trial Study protocol. Acta Oncol. 2020, 59, 336–341.
  38. Tan, L.; Sandhu, S.; Lee, R.; Li, J.; Callahan, J.; Ftouni, S.; Dhomen, N.; Middlehurst, P.; Wallace, A.; Raleigh, J.; et al. Prediction and monitoring of relapse in stage III melanoma using circulating tumor DNA. Ann. Oncol. 2019, 30, 804–814.
  39. Ricciuti, B.; Jones, G.; Severgnini, M.; Alessi, J.V.; Recondo, G.; Lawrence, M.; Forshew, T.; Lydon, C.; Nishino, M.; Cheng, M.; et al. Early plasma circulating tumor DNA (ctDNA) changes predict response to first-line pembrolizumab-based therapy in non-small cell lung cancer (NSCLC). J. Immunother. Cancer 2021, 9, e001504.
  40. Chaudhuri, A.A.; Chabon, J.J.; Lovejoy, A.F.; Newman, A.; Stehr, H.; Azad, T.; Khodadoust, M.S.; Esfahani, M.S.; Liu, C.L.; Zhou, L.; et al. Early Detection of Molecular Residual Disease in Localized Lung Cancer by Circulating Tumor DNA Profiling. Cancer Discov. 2017, 7, 1394–1403.
  41. Tarazona, N.; Gimeno-Valiente, F.; Gambardella, V.; Zuniga, S.; Rentero-Garrido, P.; Huerta, M.; Roselló, S.; Martinez-Ciarpaglini, C.; Carbonell-Asins, J.A.; Carrasco, F.; et al. Targeted next-generation sequencing of circulating-tumor DNA for tracking minimal residual disease in localized colon cancer. Ann. Oncol. 2019, 30, 1804–1812.
  42. Fitzpatrick, T.; Perrier, L.; Shakik, S.; Cairncross, Z.; Tricco, A.C.; Lix, L.; Zwarenstein, M.; Rosella, L.; Henry, D. Assessment of Long-term Follow-up of Randomized Trial Participants by Linkage to Routinely Collected Data. JAMA Netw. Open 2018, 1, e186019.
  43. Holmes, E.; Bradbury, I.; Williams, L.; Korde, L.; de Azambuja, E.; Fumagalli, D.; Moreno-Aspitia, A.; Baselga, J.; Piccart-Gebhart, M.; Dueck, A.; et al. Are we assuming too much with our statistical assumptions? Lessons learned from the ALTTO trial. Ann. Oncol. 2019, 30, 1507–1513.
  44. Jakobsen, A.; Andersen, R.F.; Hansen, T.F.; Jensen, L.H.; Faaborg, L.; Steffensen, K.D.; Thomsen, C.B.; Wen, S.W. Early ctDNA response to chemotherapy. A potential surrogate marker for overall survival. Eur. J. Cancer 2021, 149, 128–133.
More